Neurotrophic factor production promoting device

ABSTRACT

A neurotrophic factor production promoting device is provided, which is able to promote production of a neurotrophic factor or neurotrophic factor-like substance in an affected area by a simple technique that, regardless of the place of treatment, can be performed without transplantation of cells or injection into the affected area, in order to prevent or treat various diseases such as brain diseases. In order to apply a high frequency alternating magnetic field in the range of 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700 MHz to 1000 MHz to cells at a magnetic flux density of no more than 0.01 Tesla, the neurotrophic factor production promoting device includes a high frequency electromagnetic wave generating means generating a high frequency electromagnetic wave of the abovementioned frequency, in which the magnetic stimulation by the high frequency alternating electromagnetic field of the abovementioned high frequency allows the intracellular concentration of calcium ions to be increased so that exocytosis of the neurotrophic factor group is induced, and the magnetic stimulation allows messenger ribonucleic acid (mRNA) of the neurotrophic factor group to be increased in the cells so that the synthesis and extracellular release of the neurotrophic factor group are promoted.

This application is a National Stage of PCT/JP06/322295 filed Nov. 8,2006.

TECHNICAL FIELD

The present invention relates to a neurotrophic factor productionpromoting device for promoting the production of a neurotrophic factoror a neurotrophic factor-like substance, which is useful in thetreatment of various disorders such as brain diseases.

BACKGROUND ART

Brain diseases such as cerebrovascular disease, depression, andneurodegenerative disorder such as Alzheimer's disease are caused by aweakening of or damage to the central nervous system cells. In order forthese brain diseases to be treated, research is being conducted onregenerative therapies of the brain in which damaged neurocytes areprotected by transplantation of new cells into the brain or injection ofa neurotrophic factor into the brain. Although the majority of theseregenerative therapies are still under study, some are already beingapplied clinically, and thus are receiving attention as new therapeutictreatments for various brain diseases. For example, Non-Patent Documents1 and 2 disclose a therapeutic treatment for introducing a neurotrophicfactor into the brain, by transplanting cells producing a neurotrophicfactor into the brain to compensate for deficient amounts of theneurotrophic factor.

-   Non-Patent Document 1: “Regenerative Treatments of the Brain    (Nou-no-saisei-iryou),” (online), Japan Neurosurgical Society,    http://square.umin.ac.jp/neuroinf/patient/701.html (accessed Oct.    16, 2006); and-   Non-Patent Document 2: “Perspective on Neuroprotective and Neural    Repairing Agents (Shinkei-hogo, shinkei-shuhukuyaku-no-tenbou),”    (online), http://www.h2.dion ne.jp/˜park/index1/i1014hogo.html    (accessed Oct. 16, 2006).

DISCLOSURE OF THE INVENTION Problem(s) to be Solved by the Invention

Incidentally, since a neurotrophic factor having a function of repairingthe abovementioned central nervous system cells cannot pass through theblood-brain barrier (i.e., the barrier between the brain and bloodvessels for preventing the entry of a harmful substance into the brain),the neurotrophic factor cannot be administered into the brain via anintravenous injection, and the like. Consequently, the only conventionalregenerative therapies of the brain were either the abovementionedmethod of transplanting cells producing the neurotrophic factor into thebrain, or the method of directly injecting the neurotrophic factor intothe brain.

However, since a procedure for the injection of the neurotrophic factoror the transplantation of cells to the brain also presents considerablerisks, such as damage to the central nervous system cells and theintroduction of infections into the brain, the treatment method can onlybe achieved at specific advanced medical facilities, and thus in spiteof fact that the number of patients is increasing, patients have beenunable to easily receive a treatment anywhere.

Thus, the present invention was achieved in view of the abovementionedproblems, and as such, aims to provide a novel and improved neurotrophicfactor production promoting device, which is able to promote theproduction of the neurotrophic factor or neurotrophic factor-likesubstance in an affected area by a simple technique that, regardless ofthe place of treatment, can be performed without the transplantation ofcells or the injection into the affected area, in order to prevent ortreat various diseases such as brain diseases.

Means for Solving the Problem(s)

Although the mechanism of the effects of a magnetic treatment for braindiseases has not been completely elucidated, after thorough research bythe present inventors, it was found that applying a high frequencyalternating magnetic field of a predetermined frequency to specificcells (cells that are capable of producing the neurotrophic factor orneurotrophic factor-like substance, e.g., glial cells) of an affectedarea of a subject to be treated at an appropriate magnetic fieldintensity (e.g., no more than 0.01 Tesla) allows the concentration ofcalcium ions (Ca²⁺) within these cells to be increased so that anexocytotic reaction of the neurotrophic factor and/or neurotrophicfactor-like substance (hereinafter, “the neurotrophic factor and/orneurotrophic factor-like substance” may also be referred to as,“neurotrophic factor group”) is induced, and allows messengerribonucleic acid (mRNA) to be increased in the cells so that thesynthesis and release of the neurotrophic factor group is promoted,thereby allowing the production of the neurotrophic factor group to bepromoted.

Thus, the present inventors focused on the neurotrophic factor groupproduction promoting effect in such cells, and therefore, thoroughlyresearched a frequency of a high frequency alternating magnetic field tobe applied on cells by conducting various experiments and studies. As aresult, a frequency of a suitable high frequency alternating magneticfield capable of significantly enhancing the magnetic treatment effectof promoting the production of neurotrophic factor group has been found,to thereby allow the below-mentioned invention of the presentapplication to be achieved.

In order to solve the abovementioned problems, from a perspective of thepresent invention, a neurotrophic factor production promoting device isprovided for promoting the production of the neurotrophic factor or theneurotrophic factor-like substance via application of magneticstimulation to cells. This neurotrophic factor production promotingdevice includes a high frequency electromagnetic wave generating meansgenerating a high frequency electromagnetic wave of a high frequency forpromotion of production selected from the range of 20 MHz to 180 MHz,280 MHz to 600 MHz, or 700 MHz to 1000 MHz, in order to apply a highfrequency alternating magnetic field of the high frequency for thepromotion of production to the cells at a magnetic flux density of nomore than 0.01 Tesla, whereby the magnetic stimulation by the highfrequency alternating magnetic field of the high frequency for thepromotion of production allows the concentration of calcium ions (Ca²⁺)within the cells to be increased so that the exocytosis of theneurotrophic factor or the neurotrophic factor-like substance isinduced, and the magnetic stimulation allows the messenger ribonucleicacid (mRNA) of the neurotrophic factor or the neurotrophic factor-likesubstance to be increased in the cells so that the synthesis and theextracellular release of the neurotrophic factor or the neurotrophicfactor-like substance are promoted.

By generating a high frequency electromagnetic wave of the highfrequency for the promotion of production that is suitable for magnetictreatment via the above structure, it allows the abovementioned highfrequency alternating magnetic field of the high frequency for thepromotion of production to be emitted, and to be applied to cells suchas those of the affected area of a subject to be treated. The productionof the neurotrophic factor or the neurotrophic factor-like substance incells in an affected area of a subject to be treated is promoted viathis magnetic stimulation, to thereby allow cells that have beenweakened, damaged, or reduced in number by a disease to be regeneratedby the neurotrophic factor or the neurotrophic factor-like substance,and allow the abovementioned disease to be treated by a suitablemagnetic treatment. Such a magnetic treatment allows to easily treat orprevent a disease, without the transplantation of cells or the injectionto the affected area, regardless of the place of treatment.

Moreover, applying magnetic stimulation to cells capable of producingthe neurotrophic factor group via the abovementioned high frequencyalternating magnetic field within the range of 20 MHz to 180 MHz, 280MHz to 600 MHz, or 700 MHz to 1000 MHz allows for, for example, at leasta 2-fold increase in the neurite outgrowth of cells that have beenweakened and the like by a disease, when compared with an unstimulatedgroup, which thereby allows the magnetic treatment effect to beenhanced.

In addition, the abovementioned cells that are capable of producing theneurotrophic factor and/or neurotrophic factor-like substance may alsoinclude a glial cell, a neurocyte, a fibroblast, a vascular endothelialcell, an epidermal cell, a keratinocyte, an immunocyte, or a musclecell.

Moreover, the abovementioned neurotrophic factor may also include atleast one selected from a nerve growth factor (NGF), a brain-derivedneurotrophic factor (BDNF), a basic fibroblast growth factor-2 (FGF-2),and a glial cell-line derived neurotrophic factor (GDNF).

Furthermore, the neurotrophic factor-like substance may also include atleast one selected from adenosine, adenosine monophosphate (AMP), amanganese ion, genipin, lysophosphatidylethanolamine, ganglioside, andRho-kinase.

In addition, the abovementioned neurotrophic factor production promotingdevice may also be a treatment device employed to treat a disease causedby a weakening of, damage to, or a reduction in the number of thecentral nervous system cells or cerebrospinal nervous system cells.

Furthermore, the abovementioned disease may also be at least oneselected from a neurodegenerative disorder, depression, acerebrovascular disease, and a spinal cord injury.

Moreover, the abovementioned high frequency for the promotion ofproduction may be selected from the range of 60 MHz to 180 MHz, 280 MHzto 300 MHz, 450 MHz to 550 MHz, or 900 MHz to 950 MHz. Accordingly,since this allows for, for example, at least a 2.5-fold increase in theneurite outgrowth of the abovementioned cells weakened and the like by adisease, when compared with an unstimulated group, it is able to furtherenhance the magnetic treatment effect thereof.

In addition, the abovementioned high frequency for the promotion ofproduction may be selected from the range of 100 MHz to 160 MHz.Accordingly, since this allows for, for example, at least a 3.0-foldincrease in the neurite outgrowth of the abovementioned cells weakenedand the like by a disease, when compared with an unstimulated group, itis able to even further enhance the magnetic treatment effect thereof.

Furthermore, the abovementioned high frequency for the promotion ofproduction may be selected from the range of 120 MHz to 160 MHz.Accordingly, since this allows for, for example, at least a 3.5-foldincrease in the neurite outgrowth of the abovementioned cells weakenedand the like by a disease, when compared with an unstimulated group, itis able to remarkably enhance the magnetic treatment effect.

Moreover, the abovementioned high frequency electromagnetic wavegenerating means may also include a high frequency oscillation meansoutputting a high frequency electric current, and a high frequencyantenna generating the high frequency electromagnetic wave of the highfrequency for the promotion of production via the application of thehigh frequency electric current from the high frequency oscillationmeans. Accordingly, this allows the high frequency electromagnetic waveof the high frequency for the promotion of production to be suitablygenerated, so that the high frequency alternating magnetic field of thehigh frequency for the promotion of production can be appropriatelyapplied to the cells of the affected area of the subject to be treated.

In addition, the abovementioned high frequency electromagnetic wavegenerating means may also intermittently generate the high frequencyelectromagnetic wave, by repeating an on time period in which the highfrequency electromagnetic wave is generated and an off time period inwhich the high frequency electromagnetic wave is not generated, at apredetermined cycle. Accordingly, since this allows the high frequencyalternating magnetic field to be intermittently generated and applied tothe cells of the affected area of the subject to be treated, it ispossible to repeatedly switch between a state in which the highfrequency alternating magnetic field is applied to these cells and astate in which it is not applied to these cells. Thus, changes in thehigh frequency alternating magnetic field stimulation applied to thecells occur, so as to allow the magnetic treatment effect to beenhanced.

Furthermore, the abovementioned high frequency electromagnetic wavegenerating means may also intermittently generate the high frequencyelectromagnetic wave, by repeating a first on time period in which thehigh frequency electromagnetic wave is generated and a first off timeperiod in which the high frequency electromagnetic wave is notgenerated, at a cycle corresponding to 2.0±10% kHz. In addition, theabovementioned high frequency electromagnetic wave generating means mayalso intermittently generate the high frequency electromagnetic wave, byrepeating a second on time period in which the high frequencyelectromagnetic wave is generated and a second off time period in whichthe high frequency electromagnetic wave is not generated, at a cyclecorresponding to 7.8±10% Hz. Accordingly, the high frequency alternatingmagnetic field is intermittently generated, at a suitable time intervalthat the cells of the affected area of the subject to be treated areresponsive to, to be applied to the cells of the affected area.

Moreover, a low frequency electromagnetic wave generating meansgenerating a low frequency electromagnetic wave of a low frequency forthe promotion of production selected from the abovementioned range of2.0±10% kHz may also be included, in order to apply a low frequencyalternating magnetic field of the low frequency for the promotion ofproduction to the cells. Accordingly, this allows not only theabovementioned high frequency alternating magnetic field to be appliedto the cells of the affected area of the subject to be treated, but alsothe low frequency alternating magnetic field of the low frequency forthe promotion of production suitable for magnetic treatment to beapplied thereto, so that the magnetic treatment effect can be furtherenhanced.

In addition, the abovementioned low frequency electromagnetic wavegenerating means may also include a low frequency oscillation meansoutputting a low frequency electric current, and a low frequency antennagenerating the low frequency electromagnetic wave of the low frequencyfor the promotion of production via the application of the low frequencyelectric current from the low frequency oscillation means. Accordingly,this allows the low frequency electromagnetic wave of the low frequencyfor the promotion of production to be suitably generated, so that thelow frequency alternating magnetic field of the low frequency for thepromotion of production can be appropriately applied to the cells of theaffected area of the subject to be treated.

Furthermore, a rise time of the low frequency electric current appliedto the abovementioned low frequency antenna may be no more than 0.1μsec. Accordingly, since this allows a rate of change in an intensity ofthe low frequency alternating magnetic field to be increased, the cellsbecome more sensitized to the low frequency magnetic field.

Moreover, the abovementioned low frequency electromagnetic wavegenerating means may also intermittently generate the low frequencyelectromagnetic wave, by repeating an on time period in which the lowfrequency electromagnetic wave is generated and an off time period inwhich the low frequency electromagnetic wave is not generated, at apredetermined cycle. Accordingly, since this allows the low frequencyalternating magnetic field to be intermittently generated and applied tothe cells of the affected area of the subject to be treated, it ispossible to repeatedly switch between a state in which the low frequencyalternating magnetic field is applied to these cells and a state inwhich it is not applied to these cells. Thus, changes in the lowfrequency alternating magnetic field stimulation applied to the cellsoccur, so as to allow the magnetic treatment effect to be enhanced.

In addition, the abovementioned low frequency electromagnetic wavegenerating means may also intermittently generate the low frequencyelectromagnetic wave, by repeating a third on time period in which thelow frequency electromagnetic wave is generated and a third off timeperiod in which the low frequency electromagnetic wave is not generated,at a cycle corresponding to 7.8±10% Hz. Accordingly, the low frequencyalternating magnetic field is intermittently generated, at a suitabletime interval that the cells of the affected area of the subject to betreated are responsive to, to be applied to the cells.

Furthermore, the abovementioned high frequency electromagnetic wavegenerating means may also intermittently generate the high frequencyelectromagnetic wave, by repeating an on time period in which the highfrequency electromagnetic wave is generated and an off time period inwhich the high frequency electromagnetic wave is not generated, at apredetermined cycle, and the on time period of the high frequencyelectromagnetic wave and the on time period of the low frequencyelectromagnetic wave may be synchronized with each other. Accordingly,since the high frequency alternating magnetic field and the lowfrequency alternating magnetic field are repeatedly generated/notgenerated with the same timing, a time when both alternating magneticfields are applied to the cells of the affected area and a time whenboth alternating magnetic fields are not applied to the cells of anaffected area can be clearly separated. Thus, changes in the alternatingmagnetic field stimulation applied to these cells occur, so as to allowthe magnetic treatment effect to be enhanced.

Moreover, the abovementioned high frequency electromagnetic wavegenerating means may also generate the high frequency electromagneticwave of the high frequency for the promotion of production, byintermittently generating a high frequency electromagnetic wave of afrequency higher than the high frequency for the promotion ofproduction, at a cycle corresponding to the high frequency for thepromotion of production. Accordingly, by employing the high frequencyelectromagnetic wave of the high frequency as a carrier wave, theabovementioned high frequency electromagnetic wave of the high frequencyfor the promotion of production can be generated.

In addition, the high frequency electromagnetic wave of the highfrequency for the promotion of production generated by theabovementioned high frequency electromagnetic wave generating means mayalso include a higher harmonic wave occurring when a high frequencyelectromagnetic wave of less than the high frequency for the promotionof production is generated. Specifically, the high frequencyelectromagnetic wave generating means may also include anelectromagnetic wave generating means additionally generating the highfrequency electromagnetic wave of the high frequency for the promotionof production as a higher harmonic wave, when generating anelectromagnetic wave of a frequency that is an integer sub-multiple ofthe abovementioned high frequency for the promotion of production.

Effects of the Invention

According to the present invention described above, the production ofthe neurotrophic factor or neurotrophic factor-like substance in anaffected area is promoted by a simple magnetic treatment, so thatprevention or treatment of various diseases such as brain diseases canbe performed, regardless of the place of treatment, and without thetransplantation of cells or the injection into the affected area.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a perspective view showing an exterior of a magnetic treatmentdevice of a first embodiment of the present invention;

FIG. 2A is a plan view showing an example of an internal structure ofthe magnetic treatment device of the same embodiment;

FIG. 2B is a plan view showing an oscillation coil, and a plan viewshowing another example of the internal structure of the magnetictreatment device of the same embodiment;

FIG. 3 is a block diagram showing an example of a circuit structure ofthe magnetic treatment device of the same embodiment;

FIG. 4 is a waveform diagram showing waveforms of a high frequencyelectric current and a low frequency electric current applied to a highfrequency coil and a low frequency coil of the same embodiment;

FIG. 5A is an explanatory view showing a treatment aspect in which themagnetic treatment device of the same embodiment is employed;

FIG. 5B is an explanatory view showing a treatment aspect in which themagnetic treatment device of the same embodiment is employed;

FIG. 6 is a flow chart showing a mechanism of the magnetic treatmenteffect from the magnetic treatment device of the same embodiment;

FIG. 7 is a perspective view showing a structure of a magneticstimulation device employed in Experiment 1 of an Example of the presentinvention;

FIG. 8 is a graph showing experimental results of Experiment 1 of anExample of the present invention;

FIG. 9 is a graph showing experimental results of Experiment 2 of anExample of the present invention;

FIG. 10 is a graph showing experimental results of Experiment 5 of anExample of the present invention; and

FIG. 11 is a graph showing measurement results of a frequency of anelectromagnetic wave generated by the magnetic treatment device of theabovementioned embodiment.

EXPLANATION OF THE REFERENCE NUMERALS

-   10, 10A, 10B Magnetic treatment device (Neurotrophic factor    production promoting device)-   12 Housing-   16 Display portion-   18 Power source portion-   20 Control block-   21 Power source supply circuit-   22 Main control circuit-   23 Clock generating circuit-   24 High frequency wave oscillation means-   25 Low frequency wave oscillation means-   30, 30A, 30B High frequency coil-   40, 40A, 40B Low frequency coil-   50 Oscillation coil

BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the details of preferred embodiments of the presentinvention will be explained with reference to the appended drawings.Moreover, with regard to the specification and drawings of the presentapplication, the constituent elements having a substantially similarfunctional structure have been given the same reference numeral to omita duplicate description thereof.

First Embodiment

Hereinafter, a magnetic treatment device will be described as an exampleof a neurotrophic factor production promoting device of a firstembodiment of the present invention. The magnetic treatment devicepromotes the production of the neurotrophic factor and/or theneurotrophic factor-like substance (hereinafter “the neurotrophic factorand/or neurotrophic factor-like substance” will be referred to as a“neurotrophic factor group”) in cells by applying magnetic stimulationto the cells of an affected area of a human body, in order to preventand treat various diseases such as a brain diseases.

Structure of the Treatment Device

First, an external structure of a magnetic treatment device 10 of thepresent embodiment will be explained with reference to FIG. 1. Moreover,FIG. 1 is a perspective view showing the external structure of themagnetic treatment device 10 of the present embodiment.

As shown in FIG. 1, the magnetic treatment device 10 includes, forexample, a housing 12, an operating portion 14, and a display portion16.

The housing 12 is a case for internally storing each main apparatus ofthe magnetic treatment device 10, that is formed of a synthetic resin,such as a plastic and the like. This housing 12 includes a flatsubstantially rectangular parallel-piped shape (e.g., approximately 8 cmlength×6 cm width×2 cm height). However, the shape thereof is notspecifically limited to that of the above example. For example, theshape thereof can be modified into an arbitrary shape, such as asubstantially spherical shape, a substantially oval shape, asubstantially rod shape, a substantially cube shape, any other shapethat is easily grasped by a user, or the like. The user of the magnetictreatment device 10 grasps the housing 12, and an electromagnetic wave(including an alternating magnetic field) emitted from the magnetictreatment device 10 can be applied to the affected area, by bringing themagnetic treatment device 10 into immediate contact with the affectedarea or into a proximity that is within a predetermined distance withrespect to the affected area.

The operation portion 14 is a switch and the like, for turning themagnetic treatment device 10 on or off (e.g., for irradiating thealternating magnetic field). For example, the user can switch themagnetic treatment device 10 between an activated or deactivated mode bypressing the operation portion 14 each time.

Moreover, the display portion 16 is composed of a light-emitting lamp,such as a light-emitting diode (LED), and the like. This display portion16 may display the activated or deactivated mode of the magnetictreatment device 10, or a state of remaining power, or a state ofelectric charge of the below-mentioned power source portion (not shown),or the like. In the present embodiment, this display portion 16 iscomposed of two LEDs: a red LED 16 a and a green LED 16 b. This red LED16 a lights up when the remaining power of the battery and the like ofthe power source portion is above a predetermined level, and flasheswhen the remaining power of the battery is below this level, forexample. Moreover, the green LED 16 b lights up or flashes when themagnetic treatment device is activated, and turns off when the magnetictreatment device is deactivated.

However, the display portion 16 is not specifically limited to the aboveexample. For example, the display portion 16 may also be composed of aliquid crystal display device (LCD) that is capable of displayingcharacters or graphics and the like. Accordingly, the display portion 16is capable of displaying a variety of information, such as informationregarding the intensity or frequency of the electromagnetic wave(alternating magnetic field) irradiated by the magnetic treatment device10, time period that the irradiation is sustained, irradiation timing,treatment schedule, amount of battery remaining, time of day, andtemperature.

Next, the internal structure of the magnetic treatment device 10 of thepresent embodiment will be explained with reference to FIGS. 2A and 2B.Moreover, FIG. 2A is a plan view showing an example of the internalstructure of the magnetic treatment device 10 of the present embodiment(magnetic treatment device 10A); and FIG. 2B is plan view showinganother example of the internal structure of the magnetic treatmentdevice 10 of the present embodiment (magnetic treatment device 10B), anda perspective view showing an oscillation coil 50 within.

As shown in FIG. 2A, for example, a power source portion 18, a controlblock 20, a high frequency coil 30A, and a low frequency coil 40A areprovided within the housing 12 of the magnetic treatment device 10A.Among these, the control block 20, the high frequency coil 30A, and thelow frequency coil 40A, for example, are mounted on a same substrate 17,and are collectively attachable to/detachable from the housing 12.

The power source portion 18 is a direct current power source devicecomposed of a battery (e.g., 9V dry battery), such as various types ofrechargeable batteries and dry batteries, which supplies electric powerto each portion of the magnetic treatment device 10A. Moreover, thecontrol block 20 which is a circuit board equipped with a control devicecontrolling each portion in the magnetic treatment device 10A, a highfrequency oscillation circuit oscillating a high frequency wave, a clockgenerating circuit, and the like (none is shown) will be described indetail hereinafter (see FIG. 3).

The high frequency coil 30A is an example of an antenna (high frequencyantenna) emitting a high frequency electromagnetic wave by being applieda high frequency electric current. This high frequency coil 30A is aloop antenna composed of a coil having eight turns of a comparativelythick copper wire, for example. The above high frequency coil 30A is,for example, capable of generating a high frequency electromagnetic wavethat is a high frequency for the promotion of production (e.g., 100 MHzto 160 MHz) by being applied the high frequency electric current fromthe abovementioned control block 20, and emitting it peripherally. Thishigh frequency electromagnetic wave includes a high frequencyalternating magnetic field and a high frequency alternating electricfield.

On the other hand, the low frequency coil 40A is an example of anantenna (low frequency antenna) emitting a low frequency electromagneticwave by being applied a low frequency electric current. This lowfrequency coil 40A is, for example, a loop antenna composed of a coilhaving 500 turns of a comparatively thin copper wire on a shaft core.The above low frequency coil 40A is, for example, capable of generatinga low frequency electromagnetic wave, which is at a frequency ofapproximately 2.0 kHz, by being applied the low frequency electriccurrent from the abovementioned control block 20, and emitting itperipherally. This low frequency electromagnetic wave includes a lowfrequency alternating magnetic field and a low frequency alternatingelectric field.

The high frequency coil 30A and the low frequency coil 40A are mountedin parallel, for example, so as to have the central shafts thereof in,for example, substantially the same direction. Also, the central shaftsof the high frequency coil 30A and the low frequency coil 40A aredisposed so as to be parallel to the widest surface (upper and lowersurfaces of FIG. 1) of the housing 12. Thus, the high frequencyalternating magnetic field and the low frequency alternating magneticfield are formed so as to have a line of magnetic force perpendicular toa side surface of the housing 12, via the high frequency electromagneticwave and the low frequency electromagnetic wave generated by the highfrequency coil 30A and the low frequency coil 40A.

Next, the magnetic treatment device 10B shown in FIG. 2B will bedescribed. As shown in FIG. 2B, for example, a power source portion 18,a control block 20, and an oscillation coil 50 including a highfrequency coil 30B and a low frequency coil 40B are provided within thehousing 12 of the magnetic treatment device 10B. Among these, thecontrol block 20, the high frequency coil 30B, and the low frequencycoil 40B, for example, are mounted on a same substrate 17, and arecollectively attachable to/detachable from the housing 12. Moreover,when the magnetic treatment device 10B of FIG. 2B is compared with theabovementioned magnetic treatment device 10A of FIG. 2A, since only thestructure and arrangement of the high frequency coil 30B and the lowfrequency coil 40B are different therefrom, the detailed description ofall the other substantially similar constituent elements will beomitted.

As shown in FIG. 2B, the oscillation coil 50 is formed, for example, ofthe high frequency coil 30B coiled with a comparatively thick copperwire, and the low frequency coil 40B coiled with a comparatively thinconductive wire, on the outer periphery of an acrylic annular baseportion 52 having a diameter of 3 cm, an axial width of 9 mm, and aradial thickness of 2 mm. Among these, the high frequency coil 30B is asingle turn solenoid coil (diameter of 3 cm); and the low frequency coil40B is a 200 turn solenoid coil (diameter of 3 cm, and a winding widthof 5 mm). In this manner, the oscillation coil 50 is a coil having twokinds of coils, the high frequency coil 30B and the low frequency coil40B, coaxially formed on a single acrylic annular base portion 52.

This oscillation coil 50 is arranged with the central shafts thereof (ofthe high frequency coil 30B and the low frequency coil 40B) disposed soas to be perpendicular to the widest surface (upper and lower surfacesof FIG. 1) of the housing 12. Thus, the high frequency alternatingmagnetic field and the low frequency alternating magnetic field areformed so as to have a line of magnetic force perpendicular to thewidest surface of the housing 12, via the high frequency electromagneticwave and the low frequency electromagnetic wave generated by the highfrequency coil 30B and the low frequency coil 40B.

As mentioned above, the two structural examples (10A and 10B) of themagnetic treatment device 10 were described with reference to FIGS. 2Aand 2B. The abovementioned magnetic treatment device 10A and themagnetic treatment device 10B differ in their shape and a direction thatthe lines of magnetic force of the alternating magnetic fields areformed. However, in either case, the high frequency electromagnetic waveand the low frequency electromagnetic wave generated by the highfrequency coil 30A, 30B (hereinafter collectively referred to as “highfrequency coil 30”) and by the low frequency coil 40A, 40B (hereinaftercollectively referred to as “low frequency coil 40”) are irradiated, forexample, so as to be substantially uniformly dispersed in allcircumferential directions with the central shaft of the coil as acenter thereof. Thus, the magnetic treatment effect is providedregardless of which surface of the magnetic treatment device 10A, 10B isin direct contact with or in the proximity of the affected area at whichangle. Accordingly, treatment using the magnetic treatment device 10becomes easy.

Furthermore, the antenna emitting the high frequency electromagneticwave or the low frequency electromagnetic wave is not specificallylimited to the abovementioned examples of loop antenna such as the highfrequency coil 30 and the low frequency coil 40 of FIG. 2. For example,various types of antennas, such as rod antenna and the like, may beemployed.

Next, the structure and operation of the circuit of the magnetictreatment device 10 of the present embodiment will be explained with indetail reference to FIG. 3. Moreover, FIG. 3 is a block diagram showinga circuit structure of the magnetic treatment device 10 of the presentembodiment.

Moreover, the below-described control block 20 and the abovementionedhigh frequency coil 30 are one example of a structure of the highfrequency electromagnetic wave generating means generating the highfrequency electromagnetic wave of a predetermined frequency for thepromotion of production (e.g., 83.3 MHz). In addition, this controlblock 20 and the abovementioned low frequency coil 40 are one example ofthe structure of the low frequency electromagnetic wave generating meansgenerating the low frequency electromagnetic wave of a predeterminedfrequency (e.g., 2 kHz).

As shown in FIG. 3, the control block 20 includes, for example, a maincontrol circuit 22, a power source supply circuit 21, a clock generatingcircuit 23, a high frequency oscillation means 24, and a low frequencyoscillation means 25.

The main control circuit 22 is composed, for example, of a one-chipmicrocomputer, and functions to control each portion within the controlblock 20.

The power source supply circuit 21 includes, for example, an on/offcontrol circuit 212, a booster circuit 214, and a step-down circuit 216,and functions to control the supply of electric power from theabovementioned power source portion 18 to each portion within thecontrol block 20. Specifically, the on/off control circuit 212, forexample, detects the on/off of the switch of the operation portion 14,and inputs the detection results into the main control circuit 22. Inaddition, the on/off control circuit 212 turns the electric power supplyfrom the power source portion 18 to the high frequency coil 30 and thelow frequency coil 40, and the like, on or off, based on an on/offinstruction from the main control circuit 22.

Moreover, the booster circuit 214, for example, is capable of boostingthe electric power from the power source portion 18 that is composed ofa 9V dry battery, when necessary. Accordingly, the voltage supplied tothe high frequency coil 30 and the low frequency coil 40 can bemaintained at 9V, for example. Furthermore, the booster circuit 214, forexample, can also output a battery consumption error signal to the maincontrol circuit 22 when the voltage that can be output therefrom fallsbelow a predetermined level as a result of the battery consumption ofthe power source portion 18, and the like. As a result, when the errorsignal is input, the main control circuit 22, for example, controls theswitching from lightning to flashing of the red LED 16 a, so that theuser can be notified of the battery consumption.

In addition, the step-down circuit 216 is capable of maintaining thevoltage supplied to the main control circuit 22 and the like at 5V, forexample, via a step down in the power of the power source portion 18.Moreover, the step-down circuit 216, for example, can also output avoltage reduction error signal to the main control circuit 22 when thevoltage that can be output therefrom falls below a predetermined level,as a result of the battery consumption of the power source portion 18,and the like. As result thereof, the main control circuit 22 controlsthe overall operation of the magnetic treatment device 10 so as toprevent trouble, such as an unexpected stoppage in operation occurringfrom a reduction in voltage, and the like, for example. As a result,since the green LED 16 b, which is lighted during the operation of themagnetic treatment device 10, is controlled to be turned off, the usercan be notified when the operation of the magnetic treatment device 10is stopped.

The clock generating circuit 23, for example, generates a clock signalat a predetermined frequency, which is output to the main controlcircuit 22. This clock generating circuit 23, for example, isconstructed so as to be capable of generating a 32.7 kHz clock signaland a 10 MHz clock signal. The main control circuit 22 outputs a clocksignal inputted from this clock generating circuit 23 to the lowfrequency oscillation circuit 254. The low frequency oscillation circuit254, for example, generates a 2.0 kHz clock signal and a 7.81 Hz clocksignal based on the above clock signal, and outputs these to amodulation circuit 246 and a coil drive circuit 258, respectively.

The high frequency oscillation means 24 generates a high frequencyelectric current (e.g., approximately 83.3 MHz) of a predeterminedfrequency for the promotion of production, which is applied to the highfrequency coil 30. This high frequency oscillation means 24 includes,for example, a frequency control circuit 242, a high frequencyoscillation circuit 244, the modulation circuit 246, and a coil drivecircuit 248.

The frequency control circuit 242 functions to control the frequency ofthe high frequency wave generated by the high frequency oscillationcircuit 244. Specifically, this frequency control circuit 242, forexample, controls the frequency of the high frequency wave outputted bythe high frequency oscillation circuit 244, based on the high frequencywave fed back from the high frequency oscillation circuit 244, and afrequency setting signal from the main control circuit 22. As a result,the high frequency oscillation circuit 244 stably oscillates a highfrequency wave of 83.3 MHz, for example, which is capable of beingoutput to the modulation circuit 246. Moreover, so long as the highfrequency wave is a signal that is capable of transmitting apredetermined frequency, it may be either a high frequency electriccurrent or high frequency voltage. In addition, the high frequency waveof 83.3 MHz outputted by the abovementioned high frequency oscillationcircuit 244 is a substantially sinusoidal wave signal, for example.

The modulation circuit 246, for example, is capable of intermittentlyoutputting the high frequency wave of 83.3 MHz outputted from the highfrequency oscillation circuit 244, based on the clock signal inputtedfrom the low frequency oscillation circuit 254, with a two-step on/offprocess, for example.

The first step of the on/off process, for example, is a process ofpartially cutting and intermittently outputting the high frequency waveof 83.3 MHz inputted based on the 2.0 kHz clock signal. Specifically,the modulation circuit 246, for example, repeats a process in which,during a predetermined first on time period (e.g., 400 μsec), the highfrequency wave of 83.3 MHz is output as is, and then, during apredetermined first off time period (e.g., 100 μsec), it is output as asignal in which an amplitude of the abovementioned high frequency waveis cut. Accordingly, the modulation circuit 246, for example, is capableof turning on/off the 83.3 MHz high frequency wave inputted as aconstant sinusoidal wave at, for example, a cycle equivalent to 2.0 kHz,so as to intermittently oscillate the 83.3 MHz high frequency wave. Inother words, the modulation circuit 246, for example, is able to performa modulation process in which a signal showing a substantiallyrectangular wave of 2.0 kHz is output, with the 83.3 MHz high frequencywave inputted from the high frequency oscillation circuit 244 as acarrier wave.

Furthermore, the second step of the on/off process is one in which, forexample, the high frequency wave formed by the abovementioned first stepof the on/off process is further partially cut and intermittentlyoutput, based on the 7.81 Hz clock signal. Specifically, the modulationcircuit 246, for example, repeats a process in which, during apredetermined second on time period (e.g., 64 msec), the high frequencywave is output as is, and then, during the predetermined second off timeperiod (e.g., 64 msec), it is output as a signal in which the amplitudeof the abovementioned high frequency wave is cut. Accordingly, themodulation circuit 246, for example, turns on/off, at a cycle equivalentto 7.81 Hz, the 83.3 MHz high frequency wave that is intermittent at acycle equivalent to 2.0 kHz described above, and is able tointermittently oscillate a high frequency wave that is intermittent atan even larger cycle. In other words, the modulation circuit 246, forexample, is able to output a signal showing a substantially rectangularwave of 7.81 Hz, with the 83.3 MHz high frequency wave inputted from thehigh frequency oscillation circuit 244 as a carrier wave.

The high frequency wave formed by the two-step on/off process via thismodulation circuit 246 is input to the coil drive circuit 248. The coildrive circuit 248 amplifies the inputted high frequency wave with theelectric power from the power source supply circuit 21, intermittentlyoscillates the high frequency electric current with a frequency of 83.3MHz at the two cycles equivalent to 2.0 kHz and 7.81 Hz, and applies itto the high frequency coil 30. At such a time, the coil drive circuit248 controls a magnetic field intensity (magnetic flux density) of thehigh frequency electromagnetic wave generated by the high frequency coil30, by controlling the electric current value of the high frequencyelectric current applied to the high frequency coil 30, so that themagnetic field intensity of the high frequency alternating magneticfield applied to the affected area is within the range of 50 nT to 0.01T, for example. For example, when measuring the magnetic field intensityof the high frequency electromagnetic wave generated by the highfrequency coil 30B of the magnetic treatment device 10B of the presentembodiment, the magnetic field intensity was 1.3 μT, thus enabling ahigh frequency alternating magnetic field of no less than 50 μT to beapplied to the affected area, within an effective distance of 3 mm fromthe high frequency coil 30B.

On the other hand, the low frequency oscillation means 25, for example,generates a low frequency electric current of approximately 2 kHz, whichis applied to the low frequency coil 40. This low frequency oscillationmeans 25, for example, includes the low frequency oscillation circuit254, and the coil drive circuit 258.

As mentioned above, the low frequency oscillation circuit 254 generates,for example, a 2.0 kHz clock signal and a 7.81 Hz clock signal, based onthe clock signal inputted from the main control circuit 22, which areoutput to the modulation circuit 246 and the coil drive circuit 258,respectively. Moreover, the low frequency oscillation circuit 254, forexample, generates the 2.0 kHz low frequency wave as a substantiallyrectangular wave based on the above clock signal, and then, byimplementing an on/off process (every 64 msec) at a cycle that isequivalent to approximately 7.81 Hz on this low frequency, for example,to generate the 2.0 kHz low frequency wave that is intermittent at acycle equivalent to approximately 7.81 Hz. Specifically, the lowfrequency oscillation circuit 254, for example, repeats a process inwhich, during a predetermined third on time period (e.g., 64 msec), thelow frequency wave is output as is, and then, during a predeterminedthird off time period (e.g., 64 msec), it is output as a signal in whichthe amplitude of the abovementioned low frequency wave is cut.Accordingly, the low frequency oscillation circuit 254, for example, canintermittently oscillate the 2.0 kHz low frequency wave at an on/offcycle equivalent to 7.81 Hz. In addition, a circuit equivalent to theabovementioned frequency control circuit 242 and the modulation circuit246 may also be provided in front of or behind this low frequencyoscillation circuit 254.

The coil drive circuit 258 amplifies the low frequency wave inputtedfrom the low frequency oscillation circuit 254 with electric power fromthe power source supply circuit 21, intermittently oscillates a highfrequency electric current with a frequency of 2.0 kHz at a cycleequivalent to 7.81 Hz, so as to be applied to the low frequency coil 40.At such a time, the coil drive circuit 258 controls the magnetic fieldintensity (magnetic flux density) of the low frequency electromagneticwave generated by the low frequency coil 40, by controlling the electriccurrent value of the low frequency electric current applied to the lowfrequency coil 40, so that the magnetic field intensity of the lowfrequency alternating magnetic field applied to the affected area iswithin the range, for example, of 50 nT to 0.01 T. For example, whenmeasuring the magnetic field intensity of the high frequencyelectromagnetic wave generated by the low frequency coil 40B of themagnetic treatment device 10B of the present embodiment, the magneticfield intensity was 13 μT, thus enabling the low frequency alternatingmagnetic field of no less than 50 μT to be applied to the affected area,within an effective distance of 3 mm from the low frequency coil 40B.

Electromagnetic Wave Generation Timing

Hereinafter, waveforms of the high frequency electric current and thelow frequency electric current applied to the high frequency coil 30 andthe low frequency coil 40 of the present embodiment will be describedwith reference to FIG. 4. Moreover, FIG. 4 is a waveform diagram showingthe waveforms of the high frequency electric current and the lowfrequency electric current applied to the high frequency coil 30 and thelow frequency coil 40 of the present embodiment.

As shown in FIG. 4( a), the high frequency electric current of thefrequency for the production of promotion (e.g., approximately 83.3 MHz)is applied to the high frequency coil 30. This high frequency electriccurrent has, for example, an amplitude of 30 mA, and forms a symmetricalsubstantially sinusoidal wave with 0 A as the center thereof.

In addition, this high frequency electric current, for example, forms aninterrupted wave that is periodically on/off, rather than a continuouswave. Specifically, the high frequency electric current includes awaveform in which, for example, a 400 μsec first on time period (1),and, for example, a 100 μsec first off time period (2) are alternatelyrepeated, and is intermittent at a cycle corresponding to, for example,approximately 2.0 kHz. Additionally, the above high frequency electriccurrent includes a waveform in which, for example, a 64 msec second ontime period (3), and, for example, a 64 msec second off time period (4)are alternately repeated in an even larger time scale, and is alsointermittent at a cycle corresponding to, for example, approximately7.81 Hz. Furthermore, since this high frequency electric current is, forexample, an approximately 83.3 MHz high frequency wave, the rise timeand fall time thereof are extremely small, for example, no more than0.003 μsec.

Whereas, as shown in FIG. 4( b), the low frequency electric current witha frequency of approximately 2.0 kHz, for example, is applied to the lowfrequency coil 40. This low frequency electric current, for example,forms a rectangular wave (square wave) that alternates between the twovalues, 17 μA and OA, at a cycle of approximately 2.0 kHz. A time period(5) where this low frequency electric current becomes 17 μA is 400 μsec,for example, and a time period (6) where this low frequency electriccurrent becomes 0 A is 100 μsec, for example. Moreover, thesubstantially rectangular wave of this low frequency electric current iscontrolled so as to have a rise time of no more than 0.1 μsec, and afall time of, for example, no more than 1.0 μsec. In this manner, bysignificantly shortening the rise time and the fall time of the lowfrequency electric current applied to the low frequency coil 40 to nomore than 0.1 μsec and no more than 1.0 μsec, the amount of change perunit of time in the low frequency electromagnetic wave generated fromthe low frequency coil 40 via the application of this low frequencyelectric current can be increased. Thus, since the cells that are thetarget of magnetic stimulation are more sensitized (i.e., the cells arevery susceptible to magnetic stimulation) to an extremely weak magneticfield (e.g., magnetic flux density of 50 nT to 0.01 T), the productionof the neurotrophic factor group can be further promoted. Moreover,although the rate of magnetic field change generated in proximity of thecoil is not increased even when the rise time of the voltage applied tothe low frequency coil 40 is shortened, the rate of magnetic fieldchange can be increased by shortening the rise time of the low frequencyelectric current as described above, so that the cells are sensitized tothe magnetic field.

Furthermore, this low frequency electric current also forms aninterrupted wave that is periodically turned on/off at, for example,approximately 7.81 Hz, rather than a continuous wave. Specifically, thelow frequency electric current includes a waveform in which, forexample, a 64 msec third on time period (6), and, for example, a 64 msecthird off time period (8) are alternately repeated, and is intermittentat cycle corresponding to, for example, approximately 7.81 Hz.

Moreover, when comparing FIG. 4( a) and FIG. 4( b), the timing of thehigh frequency electric current being turned on/off at a cycle of 7.81Hz and the timing of the low frequency electric current being turnedon/off at a cycle of 7.81 Hz are synchronized. More specifically, thehigh frequency electric current and the low frequency electric currentare both intermittent at a cycle corresponding to 7.81 Hz (specifically,for example, repeatedly turned on/off at a cycle of 128 msec). At thistime, the timings of application of the high frequency electric currentand the low frequency electric current are controlled so that the secondon time period of the high frequency electric current (3) (or, thesecond off time period (4)) and a third on time period of the lowfrequency electric current (7) (or, the third off time period (8)) haveapproximately the same timing.

In addition, the time period in which the high frequency electriccurrent is applied to the high frequency coil 30 (1) (on time period ofthe high frequency electric current), and the time period in which thelow frequency electric current is, for example, 17 μA (5) (i.e., timeperiod in which the electric current flows to the low frequency coil 40)are synchronized. More specifically, while the high frequency electriccurrent is intermittent at 2.0 kHz (specifically, for example,repeatedly turned on/off at a cycle of 500 μsec), the low frequencyelectric current is alternated between the two values, of 17 μA and 0 A,at 2.0 kHz. In this case, the first on time period of the high frequencyelectric current (1) and the time period in which the low frequencyelectric current is 17 μA (5) are matched, and the first off time periodof the high frequency electric current (2) and the time period in whichthe low frequency electric current is 0 A (6) are the same. In thismanner, the timings of the application of the high frequency electriccurrent and the low frequency electric current are controlled so thatthe time period in which the high frequency electric current actuallyflows to the high frequency coil 30 and the time period in which the lowfrequency electric current actually flows to the low frequency coil 40are synchronized.

By applying the high frequency electric current described above at, forexample, 9V, the high frequency coil 30 is capable of generating andperipherally emitting a high frequency electromagnetic wave with awaveform that is substantially the same, for example, as that of thehigh frequency electric current shown in FIG. 4( a). This high frequencyelectromagnetic wave, for example, is a high frequency substantiallysinusoidal wave with a frequency of approximately 83.3 MHz, and isperiodically intermittent at cycles equivalent to approximately 2.0 kHzand approximately 7.81 Hz. By irradiating the above high frequencyelectromagnetic wave, for example, a high frequency alternating magneticfield of the high frequency wave for the promotion of production of 83.3MHz, for example, can be intermittently generated in the periphery ofthe magnetic treatment device 10.

More specifically, this high frequency alternating magnetic field, forexample, has a magnetic flux density (magnetic field intensity) that isperiodically increased and decreased at approximately 83.3 MHz, with amaximum amplitude of, for example, 1.3 μT, and is an alternatingmagnetic field in which the orientation of the magnetic field in both apositive and negative direction periodically fluctuates at approximately83.3 MHz, and is intermittently generated at cycles that are equivalentto, for example, approximately 2.0 kHz and approximately 7.81 Hz.

By generating the intermittent high frequency alternating magnetic fielddescribed above, the magnetic treatment device 10 is not only capable offunctioning so as to emit the approximately 83.3 MHz high frequencyalternating magnetic field that is the high frequency for the promotionof production to the subject to be treated (e.g., affected area of thehuman body, and the like), but is also capable of simultaneouslyirradiating low frequency alternating magnetic fields of approximately2.0 kHz and approximately 7.81 Hz, with the high frequency alternatingmagnetic field as a carrier wave.

Moreover, by applying the low frequency electric current described aboveat, for example, 9V, the low frequency coil 40 is capable of generatingand emitting peripherally a low frequency electromagnetic wave with awaveform that is substantially the same, for example, as that of the lowfrequency electric current shown in FIG. 4( b). This low frequencyelectromagnetic wave, for example, is a low frequency substantiallyrectangular wave with a frequency of approximately 2.0 kHz, and isperiodically intermittent at approximately 7.81 Hz. By irradiating sucha low frequency electromagnetic wave, for example, the low frequencyalternating magnetic field of a low frequency for the promotion ofproduction of approximately 2.0 kHz, for example, can be intermittentlygenerated in the periphery of the magnetic treatment device 10.

More specifically, this low frequency alternating magnetic field, forexample, has a magnetic field intensity of 13 μT, is an alternatingmagnetic field occurring by turning on/off at a cycle of 2.0 kHz, amagnetic field with a magnetic field orientation fixed, for example,solely in a positive direction (e.g., repeatedly alternating between a400 μsec on time period and a 100 μsec off time period), and overall, isintermittently generated at a cycle of approximately 7.81 Hz.

By generating the intermittent low frequency alternating magnetic fielddescribed above, the magnetic treatment device 10 is not only capable offunctioning so as to emit the approximately 2.0 kHz low frequencyalternating magnetic field with the low frequency for the promotion ofproduction on the subject to be treated, but is also capable ofsimultaneously irradiating a low frequency alternating magnetic field ofapproximately 7.81 Hz, with this low frequency alternating magneticfield as a carrier wave.

Moreover, applying the abovementioned high frequency electric currentand low frequency electric current simultaneously in parallel to thehigh frequency coil 30 and the low frequency coil 40, allows the abovehigh frequency electromagnetic wave and low frequency electromagneticwave to be generated simultaneously. As a result, for example, the highfrequency alternating magnetic field and the low frequency alternatingmagnetic field can be simultaneously generated in the periphery of themagnetic treatment device 10. At such a time, as shown by theabovementioned FIG. 4, for example, the intermittent timings, at 7.81Hz, of the high frequency electromagnetic wave and low frequencyelectromagnetic wave are mutually synchronized, and the intermittenttiming of the 2.0 kHz high frequency electromagnetic wave and themagnetic field generating timing of the 2.0 kHz low frequencyelectromagnetic wave are synchronized.

Accordingly, the timing for the generation of the high frequencyalternating magnetic field by the irradiation of the high frequencyelectromagnetic wave and the timing for the generation of the magneticfield by the irradiation of the low frequency electromagnetic wave aresynchronized. Specifically, while the low frequency coil 40 alsogenerates a magnetic field of a predetermined intensity when the highfrequency alternating magnetic field is generated by the high frequencycoil 30, the low frequency coil 40 does not also generate a magneticfield at a predetermined level when the high frequency alternatingmagnetic field is not generated by the high frequency coil 30. On thewhole, the magnetic treatment device 10 is consequently capable ofperiodically repeating the generation/non-generation of the magneticfield (the high frequency alternating magnetic field generated by thehigh frequency coil 30 and the magnetic field of a predetermined levelgenerated by the low frequency coil 40).

Moreover, although the generation of the alternating magnetic field isdescribed above, the high frequency alternating magnetic field and thelow frequency alternating magnetic field are also generated by theirradiation of the abovementioned electromagnetic wave. Since the modesof generation of these alternating magnetic fields, for example, aresubstantially the same as the mode of generation of the abovementionedalternating magnetic field, the details thereof will be omitted.

Furthermore, although examples that the 83.3 MHz high frequencyelectromagnetic wave is generated as the high frequency wave for thepromotion of production and the 2.0 kHz low frequency electromagneticwave is generated as the low frequency for the promotion of productionare described in the abovementioned FIGS. 3 and 4, the frequencygenerated is not specifically limited to the above examples. The abovemagnetic treatment device 10 of the present embodiment is capable ofgenerating a high frequency electromagnetic wave within the range of,for example, 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700 MHz to 1000MHz as a high frequency for the promotion of production with the samestructure as that above, and moreover, capable of generating a lowfrequency electromagnetic wave within the range of, for example, 2±10%kHz as a low frequency for the promotion of production.

Magnetic Treatment Mode Via the Magnetic Treatment Device

Next, the magnetic treatment mode via the above magnetic treatmentdevice 10 of the present embodiment and the effects thereof will bedescribed with reference to FIGS. 5A and 5B. Furthermore, FIGS. 5A and5B are explanatory diagrams showing the treatment mode employing theabove magnetic treatment device 10A, 10B (see FIGS. 2A and 2B) of thepresent embodiment.

As shown in FIGS. 5A and 5B, the magnetic treatment device 10 is acompact lightweight treatment device (e.g., household treatment device)operated by a battery, such as a dry battery and the like, and as such,easily portable by patients. In addition, this magnetic treatment device10 is a magnetic stimulation type treatment device that is capable ofapplying magnetic stimulation to the cells of an affected area withinthe brain and the like, from outside the body, via the generation of theelectromagnetic wave by the abovementioned high frequency coil 30 andthe low frequency coil 40. Thus, the magnetic treatment device 10 doesnot require a particular large unit, or an electrode for passingelectric current to human body as with a conventional electrode patchtype treatment device, and moreover, the head of the subject does notneed to be shaved even when the magnetic stimulation is applied to theinside of the brain, and the like.

When treating the affected area of the human body (the subject to betreated) by employing the above magnetic treatment device 10, forexample, as shown in FIGS. 5A(a) and 5B(b), the magnetic treatmentdevice 10 operated by power from the power source may be brought intodirect contact with the affected area, or only indirectly brought intocontact through hair or clothing, or the like. Thereby, the magnetictreatment device 10 is capable of applying the alternating magneticfield (the high frequency alternating magnetic field and the lowfrequency alternating magnetic field) generated in the abovementionedmanner to the cells of the affected area that are the target of magneticstimulation. At such a time, the alternating magnetic field provides themagnetic stimulation to these cells, for example, by being applied notonly to the cells on the surface of the human body (e.g., hair andskin), but also to the cells inside the human body (e.g., brain, spinal,muscle, blood vessels, and bone).

Moreover, the magnetic treatment device 10, for example, can apply theabovementioned alternating magnetic field to the cells of the affectedarea even when only brought within the proximity of a position separatedfrom the surface of the body by a predetermined distance, withoutnecessarily requiring that it be brought into direct contact with theaffected area, as shown in FIGS. 5A(b) and 5B(b). Specifically, themagnetic treatment device 10 differs, for example, from an electrodepatch type magnetic treatment device of direct contact type, and isavailable as a non-contact type magnetic treatment device that iscapable of treatment even from above the hair and clothing, and thelike. However, since the intensity of the alternating magnetic fieldgenerated by magnetic treatment device 10 decreases along with theincrease in the distance from the magnetic treatment device 10 (reducedin proportion to the cube of the distance of separation), the effect ofthe electromagnetic treatment is weakened when there is an excessiveseparation between the magnetic treatment device 10 and the affectedarea.

Thus, in view of the distance of the high frequency coil 30 and lowfrequency coil 40 from the affected area that is the target of magneticstimulation (effective distance for providing magnetic stimulation ofmagnetic field intensity of a minimal level), the magnetic treatmentdevice 10 of the present embodiment is capable of controlling themagnetic field intensity of the high frequency alternating magneticfield and the low frequency alternating magnetic field generated by thehigh frequency coil 30 and the low frequency coil 40, so as to have analternating magnetic field with a magnetic field intensity (magneticflux density) of, for example, 50 nT to 0.01 T applied to the affectedarea. The magnetic stimulation that can be provided to the cells issufficiently capable of promoting the production of the neurotrophicfactor group, even with the weak magnetic field intensity of 50 nT.Since the magnetic intensity of the earth is approximately 66 mT, theabovementioned magnetic field intensity of 50 nT is an extremely weakintensity, which is approximately 1/1000 that of the magnetism of theearth.

Hereinafter, a difference between when employing the abovementionedmagnetic treatment device 10A (see FIG. 2A) and when employing themagnetic treatment device 10B (see FIG. 2B) will be described. As shownin FIG. 5A, when the abovementioned magnetic treatment device 10A isemployed, an alternating magnetic field is generated so that the linesof magnetic force intersect in the longitudinal direction of themagnetic treatment device 10A. Moreover, as shown in FIG. 5B, when theabovementioned magnetic treatment device 10B is employed, an alternatingmagnetic field is generated so that the lines of magnetic forceintersect in the lateral direction of the magnetic treatment device 10B.Accordingly, the magnetic stimulation is capable of deeply and broadlypenetrating into the brain of a patient. Moreover, since the magnetictreatment device 10A, 10B generates the alternating magnetic fieldtowards the overall periphery thereof, having the orientation of themagnetic treatment device 10A, 10B oppose the affected area at the timeof magnetic treatment allows for a treatment in which the alternatingmagnetic field is applied to the affected area even in an arbitrarydirection, such as a direction that is perpendicular to or oblique tothe surface of the body, without being specifically limited to theexample in which the direction thereof is parallel to the surface of thebody.

By applying the high frequency alternating magnetic field and the lowfrequency alternating magnetic field to the affected area using theabovementioned magnetic treatment device 10 in the above-indicatedmanner, for example, the production of the neurotrophic factor group inspecific cells of the affected area is promoted, and the recovery ofcentral nervous system cells or cranial nerve cells is stimulated by theneurotrophic factor group, so that an magnetic treatment effect can beproduced whereby brain diseases and the like are treated.

At such a time, since the magnetic treatment device 10 intermittentlyoscillates the abovementioned high frequency electromagnetic wave andthe low frequency electromagnetic wave, the alternating magnetic fieldis intermittently applied to the affected area, so that the effect ofthe magnetic field can be altered. Thus, the magnetic treatment effectis not weakened due to the tissue of an affected area (cells, and thelike) becoming accustomed to a constant magnetic field, such as in acase where the constant alternating magnetic field is continuouslygenerated. Moreover, since the timing for the generation of the highfrequency alternating magnetic field and the timing for the generationof the low frequency alternating magnetic field are synchronized, theoverall magnetic field applied by the magnetic treatment device 10 has adistinct on/off pattern. Thus, the presence or absence of magneticstimulation to the affected area is further clarified, so that themagnetic treatment effect can be enhanced.

Furthermore, a rise time and fall time of the magnetic treatment device10 are extremely small at, for example, no more than 0.003 μsec for thegenerated high frequency electromagnetic wave, and a rise time and falltime of the low frequency electromagnetic wave that is substantiallyrectangular are adjusted to no more than 0.1 μsec and no more than 1.0μsec, for example. Thus, during a change in the abovementionedalternating magnetic field, the rate of change to/from theapplication/non-application of the magnetic field is rapid. Accordingly,since the tissue of the affected area is sensitized to a change in thismagnetic field, the magnetic treatment effect is enhanced.

In addition, the magnetic treatment device 10 provides a feature inwhich the intensity (magnetic flux density) of the high frequencyalternating magnetic field and the low frequency alternating magneticfield applied to the cells of the affected area is, for example, no lessthan 50 nT to no more than 0.01 T, which is extremely small whencompared with that of another conventional magnetic treatment device(e.g., 0.8 T to 10 T). Specifically, when a magnetic field of a highintensity such as that of a conventional magnetic treatment device isemployed, damage to the affected area of the brain and the like mayoccur from the magnetic stimulation. Since damage from such a strongmagnetic stimulation is well-known, the safety operation guidelines fora magnetic field environment have been established by industrializedcountries. For example, in America (Standford University, 1971), thedaily exposure to magnetic stimulation to be applied to a region of thebody or the head is made 0.02 T and several minutes per day.

However, a large majority of conventional magnetic treatment methods ormagnetic stimulation methods constitute those applying a magnetic fieldof at least 0.1 T. For example, even the magnetic field intensity of themagnet for the treatment of shoulder stiffness with a diameter ofseveral millimeters is 0.08 T to 0.13 T. As described above, it isundeniable that a conventional magnetic treatment device employingmagnetic stimulation via a high intensity magnetic field may causedamage to a living body when used for an extended period of time.Whereas, since the magnetic field intensity applied to the cells of theaffected area by the magnetic treatment device 10 of the presentembodiment is within the extremely weak range of no less than 50 nT tono more than 0.01 T described above, the possibility of damage beingcaused to a living body is extremely low. Specifically, a safe magnetictreatment can be provided to an affected area that is important andsensitive, such as the brain.

Nonetheless, it is thought that when the magnetic field intensity of thealternating magnetic field applied to the affected area is excessivelylow (e.g., less than 30 nT), the magnetic treatment effect thereof isreduced. One theory is that the minimal magnetic field intensity thatthe cells are capable of reacting to is about 30 nT, for example. Insuch a case, in order for the magnetic treatment device 10 of thepresent embodiment to have a magnetic field intensity that is in thepreferable range of, for example, 50 nT to 0.01 T applied to the cellsof the affected area, the magnetic field intensity of the high frequencyalternating magnetic field and the low frequency alternating magneticfield generated by the high frequency coil 30 and the low frequency coil40 is controlled. The magnetic field intensity of the alternatingmagnetic fields generated by these coils is determined based on thedistance between the affected area which is the target of magneticstimulation and the high frequency coil 30 as well as the low frequencycoil 40 within the magnetic treatment device 10 (e.g., the distance fromthe surface of the body to the affected area within the brain), themagnetic permeability of the affected area (e.g., the magneticpermeability of the brain), and the like.

Specifically, with regard to the structure of the magnetic treatmentdevice 10B of the present embodiment, when a high frequency alternatingmagnetic field and a low frequency alternating magnetic field of atleast 50 nT are applied to an affected area with a depth of, forexample, no more than 6 cm from the surface of the body (specifically,when the effective distance of magnetic stimulation by the magnetictreatment device 10 is 6 cm), the magnetic field intensity of the highfrequency alternating magnetic field generated by the high frequencycoil 30B in the proximity of the above coil 30B is set at, for example,approximately 0.01 T, and the magnetic field intensity of the lowfrequency alternating magnetic field generated by the low frequency coil40B in the proximity of the above coil 40B is set at, for example, noless than approximately 0.1 T. Moreover, when the abovementionedeffective distance is 12 cm, the magnetic field intensity of theabovementioned high frequency alternating magnetic field in theproximity of the coil 30B is set at, for example, approximately 0.1 T,and the magnetic field intensity of the abovementioned low frequencyalternating magnetic field in the proximity of the coil 40B is set at,for example, approximately 1 T.

As described above, for example, the magnetic treatment device 10 of thepresent embodiment is capable of applying an alternating magnetic fieldof a preferred frequency and magnetic field intensity to promote theproduction of neurotrophic factor group of the cells of the affectedarea, and of switching between the application/non-application of theabove alternating magnetic field at a timing in which the cells areeasily susceptible to stimulation. Accordingly, the magnetic treatmentdevice 10 of the present embodiment demonstrates an extremely effectivemagnetic treatment effect when compared to a conventional magnetictreatment device.

Furthermore, not only is the magnetic treatment device 10battery-powered, light, compact and easily portable in addition to beingeasy to operate, it is also capable of easily and quickly (e.g., 10minutes) achieving the magnetic treatment effect by being brought indirect contact with or into the proximity of the affected area asdescribed above. Accordingly, an magnetic treatment employing the abovemagnetic treatment device 10 does not require advanced medicaltechnology, such as a cell transplantation to the inside of the brain orinjection into the brain, like a conventional regenerative therapy forthe brain. Thus, the patient himself/herself is able to use the magnetictreatment device 10 at an arbitrary location, such as home, work, orschool, without being admitted to a hospital, so that the treatment canbe easily performed at anytime.

In addition, a conventional regenerative therapy, such as a conventionalcell transplantation to the inside of the brain or injection into thebrain, may lead to brain damage, an infectious disease thereof, or sideeffects. Whereas, by applying magnetic stimulation to cells functioningto produce neurotrophic factor group from outside the body, the magnetictreatment device 10 of the present embodiment promotes the production ofneurotrophic factor group within the above cells and stimulates theindependent recovery or proliferation of weakened central nervous systemcells or cerebrospinal nervous system cells. Thus, since the insertionof a medical apparatus within the brain is not required, as with a celltransplantation or an injection, it has a significant advantage in whichthere is no brain damage, infectious diseases thereof, or side effects,and that the effect on the cells or tissue around the affected area issmall.

Therapeutic Target of the Magnetic Treatment Device

Next, with respect to the magnetic treatment device 10 of the presentembodiment, (1) the cells that are the target of magnetic stimulation,(2) the substances produced via the magnetic stimulation, (3) the sitethat is the target of treatment, and (4) the disease that is the targetof treatment, will be described in detail.

(1) Cells that are Target of Magnetic Stimulation (Cells that areCapable of Producing the Neurotrophic Factor Group)

The cells that are the target of magnetic stimulation via the magnetictreatment device 10 are cells that are capable of producing theneurotrophic factor and/or the neurotrophic factor-like substance.Specifically, the cells that are the target of this magnetic stimulationare, for example, glial cells, neurocytes, fibroblasts, vascularendothelial cells, muscle cells, epidermal cells, keratinocytes,immunocytes, and the like. The primary sites where the cells that arethe target of this type of magnetic stimulation exist are, for example,the brain, the spinal cord, nerves, blood vessels, muscles, skin, andthe like.

Among these, glial cells (neuroglia) is a generic term of therepresentative cells producing the neurotrophic factor group, and, forexample, there are astrocytes (astroglia), microcytes (microglia),oligodendrocytes, Schwann cells, mantle cells, and the like. Glial cellsare present, for example, in the brain, in the periphery of nerve cells,in blood vessels, in muscle, and the like, and the neurotrophic factorgroup that is produced by the glial cells themselves is supplied to theglial cells or neurocytes, in order to aid in the recovery andproliferation of these cells. Moreover, neurotrophic factor group isalso produced by non-nerve related cells present in areas throughout thebody, such as the abovementioned fibroblasts, vascular endothelialcells, muscle cells, epidermal cells, immunocytes, and the like.

(2) Substances Produced Via Magnetic Stimulation

When the magnetic stimulation is applied to the above cells that are thetarget of magnetic stimulation (e.g., glial cells) via the magnetictreatment device 10, the neurotrophic factor and/or neurotrophicfactor-like substance is produced in these cells. The physiologicaleffect of this neurotrophic factor and/or neurotrophic factor-likesubstance is the principal magnetic treatment effect from the magnetictreatment device 10.

The neurotrophic factor (neurotrophin (NT)) is a molecule (protein)supporting the maintenance of the normal function or the survival of theneural cells that are present in the brain, the spinal cord, andperipheral nerves, and which plays an important role in the maintenance,survival, or regeneration of damaged neural cells, or thedifferentiation or growth of neural cells during a developmental period.This neurotrophic factor may include, for example, a nerve growth factor(NGF), a brain-derived neurotrophic factor (BDNF), a fibroblast growthfactor-2 (FGF-2), a glial cell line-derived neurotrophic factor (GDNF),and the like.

The neurotrophic factor-like substance is a substance group other than aneurotrophic factor that promotes neurite outgrowth in neuronal cells.This neurotrophic factor-like substance is a substance supporting themaintenance of the normal function or the survival of the neural cellsin a manner similar to that of the abovementioned neurotrophic factor,and has a proteinaceous component and a non-proteinaceous component. Forexample, the neurotrophic factor-like substance may include, adenosine,adenosine monophosphate (AMP), a manganese ion, genipin (herbal medicinederived low molecular weight substance with plant component),lysophosphatidylethanolamine (animal-plant membrane component),ganglioside, Rho-kinase, and the like. Among these, adenosine andadenosine monophosphate are non-proteinaceous neurotrophic factor-likesubstances, and Rho-kinase is a proteinaceous neurotrophic factor-likesubstance.

(3) Site that is Target of Treatment

The therapeutic target site (affected area) of the subject to be treatedis the central nervous system (CNS) or the craniospinal nervous system.The central nervous system includes: the telencephalon, thediencephalon, the mesencephalon, the cerebellum, the pons, the medullaoblongata, the spinal cord and blood vessels. The above central nervoussystem is made up of neurocytes (neurons), glial cells, and bloodvessels. Moreover, the craniospinal nervous system is a nerve systemcomposed of cranial nerves and spinal nerves that are part of theperipheral nervous system (PNS). The above craniospinal nervous systemis composed of neurocytes (neurons), Schwann cells, and mantle cells.The cells forming the above central nervous system and craniospinalnervous system undergo repair, growth, differentiation, andproliferation via the physiological effect of the neurotrophic factorgroup supplied from the abovementioned glial cells and the like, tocontribute to the treatment of the various diseases indicated below.Moreover, the differentiation of cells is the change in the property andmorphology of cells. Furthermore, central nervous system cells are cellsthat are present in the central nervous system (the telencephalon(cerebral hemisphere), the diencephalon, the mesencephalon, thecerebellum, the pons, the medulla oblongata, the spinal cord and bloodvessels). In addition, craniospinal nervous system cells are cells thatare present in the craniospinal nervous system.

Moreover, in the subject to be treated (e.g., human body), the site thatis the target of the treatment described above and the site that is thetarget of the magnetic stimulation via the magnetic treatment device 10may be the same site or may be a different site. For example, themagnetic stimulation may also be applied to the brain (the site that isthe target of magnetic stimulation) in order to treat the brain (thesite that is the target of treatment). In addition, the magneticstimulation may also be applied to another site capable of supplying theneurotrophic factor group to the spinal cord in order to treat thespinal cord (the site that is the target of treatment), for example, thefemoral region (the site that is the target of magnetic stimulation).

(4) Disease that is Target of Treatment

The disease that is the target of the treatment via the magnetictreatment device 10 is a disease caused by the weakening of, damage to,or a reduction in the number of cells forming the abovementioned centralnervous system or craniospinal nervous system (e.g., neurocytes or glialcells), as a result of various factors. Specifically, the disease thatis the target of the treatment, for example, is: (a) a neurodegenerativedisorder (e.g., Alzheimer's disease, Parkinson's disease, Huntington'sdisease, amyotrophic lateral sclerosis, multiple sclerosis, multiplesystem atrophy, and spinocerebellar degeneration); (b) depression; (c) acerebrovascular disease (e.g., stroke, and cerebral infarction); (d)chronic pain; (e) neuropathic pain; (f) a spinal cord injury (occurringfrom an external injury or lesion); and the like. Furthermore, althoughnot for a disease, one of the effects of the magnetic treatment via themagnetic treatment device 10 is also, (g) a neuroprotective effect, toprevent these diseases.

Accordingly, the magnetic treatment device 10 of the present embodimentmay be available as a neurodegenerative disorder treatment device (e.g.,Alzheimer's disease treatment device, Parkinson's disease treatmentdevice, Huntington's disease treatment device, amyotrophic lateralsclerosis treatment device, multiple sclerosis treatment device,multiple system atrophy treatment device, spinocerebellar degenerationtreatment device, and the like), a depression treatment device, acerebrovascular disease treatment device (e.g., stroke treatment device,or cerebral infarction treatment device), a chronic pain treatmentdevice, a neuropathic pain treatment device, a spinal cord injurytreatment device, or a device for the prevention of the abovementionedvarious types of diseases.

For example, when treating depression, an alternating magnetic field isirradiated on the brain of the human body using the abovementionedmagnetic treatment device 10, the secretion of BDNF, NGF or the like,from glial cells (e.g., astrocytes) in the brain is promoted, and theneurotrophic factor group is provided to central nervous system cellsthat have been weakened and the like, so that cellular function isrestored by the proliferation or regeneration of peripheral centralnervous system cells, and the production of serotonin (endorphin in thebrain) is restored, to thereby allow it to contribute to the treatmentof depression.

Moreover, when treating Alzheimer's disease, an alternating magneticfield is irradiated on the brain of the human subject using theabovementioned magnetic treatment device 10, the secretion of BDNF, NGFor the like, from glial cells (e.g., astrocytes) of the nucleus basalisof Meynert in the brain where weakening and the like has occurred ispromoted, and cerebral cortical cells that have been weakened byβ-amyloid deposits are proliferated or regenerated, to thereby allow itto contribute to the treatment of Alzheimer's disease.

Furthermore, when treating a stroke, an alternating magnetic field isirradiated on the brain of the human body using the abovementionedmagnetic treatment device 10, the secretion of BDNF, NGF or the like,from glial cells (e.g., astrocytes) of a site that has been damaged by avascular occlusion and the like is promoted, and the neurotrophic factorgroup is provided to the damaged cells, so that the neurocytes and glialcells of the damaged area are proliferated and regenerated, to therebyallow it to contribute to the treatment of a stroke.

In addition, when treating neuropathic pain, an alternating magneticfield is irradiated on an affected area in which chronic pain is felt,using the above-mentioned magnetic treatment device 10, and theproduction of BDNF, NGF or the like, is promoted at a peripheral nerve.The above BDNF, NGF or the like, moves inside neurocytes, is transportedto dorsal root ganglia or the spinal cord, and the astrocytes areproliferated in the dorsal root ganglia and the spinal cord, to allowthe recovery from nervous erethism and allow neuropathic pain to betreated. The above NGF or the like moves inside neurocytes or to aperipheral site at which it is produced, is transported to dorsal rootganglia or to the spinal cord, and repairs or regenerates damagedsensory neurons, to thereby allow it to contribute to the treatmentthereof.

Mechanism of the Magnetic Treatment Effect

Next, the mechanism for achieving the magnetic treatment effect for theabovementioned diseases via magnetic stimulation of the alternatingmagnetic field from the magnetic treatment device 10 of the presentembodiment will be described with reference to FIG. 6. FIG. 6 is aflowchart showing the mechanism of the magnetic treatment effect fromthe magnetic treatment device 10 of the present embodiment.

In a large sense, the mechanism of the magnetic treatment effect is thata high concentration of the neurotrophic factor and/or the neurotrophicfactor-like substance (hereinafter referred to as “neurotrophic factorgroup”) is supplied to central nervous system cells of an area affectedwith brain disease (e.g. brain), and the regeneration of central nervoussystem cells is promoted. Specifically, a high frequency alternatingmagnetic field is irradiated from the magnetic treatment device 10, andmagnetic stimulation is applied to cells (the cells that are the targetof the abovementioned magnetic stimulation) that are capable ofproducing the neurotrophic factor group, such as glial cells and thelike, so that the production of intracellular neurotrophic factor groupis promoted, the neurotrophic factor group produced by these cells aresupplied to the cells of an area affected with brain disease, and thelike, and the central nervous system cells that have been weakened,damaged, or reduced in number or the like, by a brain disease and thelike, are repaired, grown, differentiated, or proliferated.

As shown in FIG. 6, based on the results of cellular experiments, themechanism of the promotion of production of the neurotrophic factorgroup is thought to be the simultaneous occurrence of threeintracellular processes. These three processes are: (1) the release ofthe neurotrophic factor group via exocytosis due to an increase in theintracellular concentration of calcium ions (S10→S20→S30); (2) thesynthesis and release of neurotrophic factor group via an increase inmRNA due to an increase in the intracellular concentration of calciumions (S10→S20→S40→S42→S44); and (3) the synthesis and release ofneurotrophic factor group via an increase in mRNA that is not due to anincrease in the intracellular concentration of calcium ions(S10→S40→S42→S44). Hereinafter, each of these neurotrophic factor groupproduction processes will be described.

(1) Exocytosis

Exocytosis is a cellular function that eukaryotic cells possess, andwhich functions in the extracellular release of an intracellularlystored substance. Exocytosis also allows the extracellular release ofmacromolecules, such as proteins.

The process (1) will be described in greater detail. First, with respectto a cell producing neurotrophic factor group, such as glial cells, andthe like, magnetic stimulation (S10) from the magnetic treatment device10 allows a voltage-dependent calcium ion channel or avoltage-independent calcium ion channel that is intracellularly locatedor located on the surface of a cell membrane to be opened, allowscalcium ions from an extracellular or intracellular calcium storage siteto be supplied, and allows the intracellular concentration of calciumions to be increased (S20). The increase in the intracellularconcentration of calcium ions allows vacuoles in which intracellularneurotrophic factor group is stored to fuse with a cell membrane, andexocytosis that releases neurotrophic factor group into extracellularspaces to occur (S30). Exocytosis is induced even when the increase inthe intracellular concentration of calcium ions is a mere 10%. Therelease phenomena is started and finished within a few minutes of theincrease in the intracellular concentration of calcium ions.

(2) Increase in mRNA Associated with Increase in IntracellularConcentration of Calcium Ions

The process (2) is a process arising from the cellular proliferationpromoting effect. It is already known in the field of neurology that theneurotrophic factor group is produced even during the cell divisionstage. The abovementioned magnetic stimulation (S10) allows theintracellular concentration of calcium ions to be increased (S20), tothereby accelerate a cellular growth cycle. The cells in the quiescentstage of the cellular growth cycle pass through the G1 stage to moveinto the S stage, via the abovementioned increase in the calcium ionconcentration. During the S stage, DNA and RNA replication occurs, andmRNA for the production of the neurotrophic factor group is increased(S40: for data demonstrating the evidence, refer to the results of thebelow-mentioned Experiment 2). The mRNA increased via the abovereplication allows a neurotrophic factor protein and a partiallyproteinaceous neurotrophic factor-like substance to be synthesized(S42), and the synthesized neurotrophic factor group to beextracellularly released (S44). The production of the neurotrophicfactor group via the process (2) is thought to intensify with time afterthe magnetic stimulation. It is thought that this is validated by themechanism of cell division.

(3) Increase in mRNA that is not Due to Increase in IntracellularConcentration of Calcium Ions

During the process (3), within the cells receiving the abovementionedmagnetic stimulation (S10), mRNA is increased (S40) and neurotrophicfactor group is synthesized (S42) and released extracellulary (S44),without being associated with the increase in the intracellularconcentration of calcium ions.

Evidence that supports the assumption that the process (3) is occurringwill be described. In the below-mentioned Experiment 1, a medium of MB8cells three hours after the magnetic stimulation (the cells that are thetarget of magnetic stimulation) was added to PC12 cells (cells that aredifferentiated by the supply of neurotrophic factor group), and theneurite outgrowth of the PC12 cells was confirmed. The division of MB8cells employed in Experiment 1 occurs approximately once per day. Sincethe number of cells moving into the S stage during cultivation threehours after magnetic stimulation under experimental conditions is small,it is thought that the mRNA elevating effect is directly activated bymagnetic stimulation, as in the abovementioned process (3). Moreover, inanother experiment, experimental data were obtained which showed thatthe MAP kinase activity within neurocytes increased by approximately 20%after magnetic stimulation was conducted for 10 minutes on culturedneurocytes which were then allowed to stand for an additional 10minutes. MAP kinase is an enzyme that regulates the activation ofproteins within cells, and the synthesis of DNA and RNA in a chainreaction. This chain reaction is referred to as the mitogen-activatedprotein kinase pathway, and is a reaction system in which a signal istransmitted from the cell membrane to the nucleus. Since the synthesisof a neurotrophic factor protein is promoted when this reaction systemis activated, mRNA is also increased.

The abovementioned process (2) is one in which RNA synthesis occurs as aresult of an increase in the intracellular concentration of calciumions. However, in the abovementioned experiment, mRNA is synthesized ina reaction time of 3 hours, and as mandated thereby, the amount of theneurotrophic factor group synthesized is low. Accordingly, in view ofsuch experimental results, it is believed that the process (3) proceedssimultaneously along with the abovementioned process (2).

Next, the mechanism for the production of a non-proteinaceousneurotrophic factor-like substance will be described. Representatives ofthe neurotrophic factor-like substance include, for example, adenosine,adenosine monophosphate (AMP), a manganese ion, genipin (herbal medicinederived low molecular weight substance with plant component),lysophosphatidylethanolamine (animal-plant membrane component),ganglioside, and Rho-kinase. Although hundreds of types of neurotrophicfactor-like substances have been discovered in addition thereto, many ofthese have not been identified as substances.

These neurotrophic factor-like substances, such as a single ion (e.g.,manganese ion), a low molecular weight substance (e.g., adenosine oradenosine monophosphate), a lipid (e.g., lysophosphatidylethanolamine,or a ganglioside), or a proteinaceous component (e.g., Rho-kinase),include a wide variety of properties and types thereof. The synthesis ofproteinaceous neurotrophic factor-like substance is promoted under thedirection of mRNA, and the proteinaceous neurotrophic factor-likesubstance is released extracellularly. Whereas, there are many types ofnon-proteinaceous neurotrophic factor-like substances, and thus theproduction mechanisms thereof also vary. For example, a neurotrophicfactor-like substance of a single ion or low molecular weight substancemay be one that exists intracellularly, or may be synthesizedintracellularly. In addition, a lipid neurotrophic factor-like substanceis synthesized intracellularly. It is thought that both of thesenon-proteinaceous neurotrophic factor-like substances are releasedextracelluarly via excytosis. Since the lipid neurotrophic factor-likesubstance is also a structural component of cell membranes, it isthought that this substance may also be released extracellularly via aprocess other than exocytosis.

When the neurotrophic factor group is produced in cells (e.g., glialcells) that have received magnetic stimulation via the abovementionedprocesses (1) to (3), this neurotrophic factor group is supplied to thecentral nervous system cells and the like that are weakened by a diseaseand the like, and the central nervous system cells are provided with aprotective effect via the neurotrophic factor group (S50). As a result,the central nervous system cells that are weakened and the like areactivated and repaired, grown, differentiated, or proliferated (S60), sothat a therapeutic effect for a brain disease (e.g., a neurodegenerativedisorder, depression, or cerebrovascular disease) caused by theabove-mentioned weakening of central nervous system cells and the likecan be obtained (S70).

According to the above described mechanism, suitable magneticstimulation is applied to an affected area using the magnetic treatmentdevice 10, so that the production of the neurotrophic factor groupwithin the cells is promoted, to thereby allow a superior treatment orpreventive effect to be exerted on various diseases, such as theabovementioned brain diseases and the like.

Based on such a perspective, the magnetic treatment device 10 of thepresent embodiment is capable of emitting, to be applied to an affectedarea, an approximately 120 MHz to 160 MHz high frequency alternatingmagnetic field at a frequency for the production of promotion andapproximately 2.0 kHz low frequency alternating magnetic field, with amagnetic flux density of no more than 0.01 T, for example, as analternating magnetic field capable of providing the desired magneticstimulation. It is thought that the stimulation by the irradiation ofthis high frequency alternating magnetic field of approximately 120 MHzto 160 MHz is highly effective in promoting the production of theneurotrophic factor group in cells, when compared to, for example, otherfrequency bands. Furthermore, it is thought that the stimulation by theirradiation of the 2.0 kHz low frequency alternating magnetic field, forexample, functions to release β-endorphin or cytokine or the like fromcells.

Moreover, although the frequency for the production of promotion of thehigh frequency alternating magnetic field applied to the affected areais preferably approximately 120 MHz to 160 MHz in terms of the magnetictreatment effect according to the below-mentioned experimental results,it was found that the contribution to the increase in the intracellularconcentration of calcium ions was sufficient even with frequenciesoutside of this range. The desired range for the frequency for thepromotion of production is 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700MHz to 1000 MHz (fourth most preferable range); more preferably 60 MHzto 180 MHz, 280 MHz to 300 MHz, 450 MHz to 550 MHz, or 900 MHz to 950MHz (third most preferable range); even more preferably 100 MHz to 160MHz (second most preferable range); and most preferably 120 MHz to 160MHz (first most preferable range). Among these examples, since afrequency for the production of promotion around the latter rangeproduces even more of the neurotrophic factor group within the cellsthat are the target of the magnetic stimulation and can supply theneurotrophic factor group to cells of a site that is the target oftreatment, the magnetic treatment effect therefrom is significant.

EXAMPLES

Next, the results of experiments that were conducted in order to verifythe magnetic treatment effect via the magnetic treatment device 10 ofthe abovementioned embodiment will be described. As mentioned above,this magnetic treatment device 10 is capable of applying magneticstimulation on the subject to be treated, by emitting a high frequencyalternating magnetic field and a low frequency alternating magneticfield (e.g., 2.0 kHz) at a desired frequency for the promotion ofproduction. Moreover, the below-mentioned examples are to experimentallyverify the magnetic treatment effect of the magnetic treatment device 10of the abovementioned embodiment, and as such, the present invention isnot specifically limited to the below-mentioned examples.

Experiment 1

First, Experiment 1, which was conducted in order to determine asuitable range for a high frequency for the promotion of production forthe high frequency alternating magnetic field applied to the cells ofthe subject to be treated via the abovementioned magnetic treatmentdevice 10, will be described.

In the Experiment 1, a high frequency alternating magnetic field of aplurality of different frequencies (20 MHz to 3000 MHz) was applied tothe cells that were the target of magnetic stimulation (MB8 cells), theneurotrophic factor group was produced in a culture medium, and then thedegree of neurite outgrowth was determined by adding the culture mediumcontaining this neurotrophic factor group to PC12 cells (cells thatdifferentiate with neurites being outgrown (differentiated) via thepresence of neurotrophic factor group), to determine the effectivenessof the magnetic treatment of each frequency when compared to anunstimulated group in which no magnetic stimulation was applied.

First, the experimental conditions of the present Experiment 1 and theprocedures (1) to (5) thereof will be described.

(1) Cultivation of MB8 Cells and PC12 Cells

Glial lineage “MB8 cells” were employed as the cells producing theneurotrophic factor group (the cells that were the target of magneticstimulation). These MB8 cells are the cells that produce a neurotrophicfactor and a neurotrophic factor-like substance. The brain cells of aneight day old mouse were cultured, and glial cells were proliferated, toobtain the MB8 cells. The MB8 cells were plated into a 24-well cultureplate (collagen I coated plate) at approximately 15.5×10⁵ cells perwell, and cultured for 24 hours in a 5% carbon dioxide environment at37° C. using a 10% FBS supplemented DMEM culture medium (Manufactured byNissui Pharmaceutical Co., Ltd.) and a carbon dioxide culture apparatus(incubator).

Moreover, PC12 cells (JCRB0266) were employed as cells for theconfirmation of neurite outgrowth. These PC12 cells are adrenalmedullary pheochromocytoma cells that are normally employed in nervegrowth factor experiments and research. These PC12 cells are the cellsin which neurites are outgrown by the presence of nerve growth factor(NGF) and the like, and which initiate the differentiation of neurocytesfrom adrenal cells via the action of nerve growth factor and the like.Cells that were purchased from the Health Science Research ResourcesBank, which is a cell bank, were employed as the PC12 cells. These PC12cells were plated into a 48-well culture plate (collagen IV coatedplate) at approximately 28×10² cells per well (so that the spacesbetween two cells were not too close), and cultured for 24 hours in a 5%carbon dioxide environment at 37° C. using 10% horse serum and 5% FBSsupplemented RPMI-1640 culture medium (Manufactured by NissuiPharmaceutical Co., Ltd.) and a carbon dioxide incubator. In addition,the below-mentioned cell cultures were all conducted in a 5% carbondioxide environment at 37° C.

(2) Magnetic Stimulation on MB8 Cells

Using an experimental magnetic stimulation device equivalent to theabovementioned magnetic treatment device 10B (refer to FIG. 2B),magnetic stimulation was applied to each of the MB8 cells (cells thatproduce the neurotrophic factor group) in the above culture plate. Themagnetic stimulation was conducted from the lower surface side of theculture plate by using the magnetic stimulation device to irradiate analternating magnetic field. At such a time, it was incubated for 30minutes (no magnetic stimulation was applied during the incubationthereof) after having the magnetic stimulation applied for 30 minutes,and then the magnetic stimulation was re-applied for 30 minutes. Thefrequency of the high frequency alternating magnetic field applied tothe MB8 cells in such a manner was incrementally changed at eachexperimental unit within the range of 20 MHz to 3000 MHz, and eachexperiment was conducted.

The structure of the experimental magnetic stimulation device employingthis magnetic stimulation will be described. This magnetic stimulationdevice is composed of: a signal generating device (“E4421B”,manufactured by Agilent Technologies) for generating a high frequencywave in the MHz frequency band (20 MHz to 3000 MHz); a functiongenerator (“33220A”, manufactured by Agilent Technologies) forgenerating a low frequency wave in the kHz frequency band (2.0 kHz); afunction generator (“FG320”, manufactured by Yokogawa Electric Co.,Ltd.) for generating a low frequency wave in the Hz frequency band (7.81Hz); an RF-AMP unit (amplifier) regulating output intensity of thesignals of these three frequency bands; a control unit integrallycontrolling the signals of these three frequency bands; and theoscillation coil 50 provided in the abovementioned magnetic treatmentdevice 10B of FIG. 2B.

During magnetic stimulation, as shown in FIG. 7, the abovementioned MB8cell culture plate 60 was mounted on the oscillation coil 50, and alight shielding cloth was placed thereon. Next, a high frequencyelectric current and a low frequency electric current were respectivelyapplied to a high frequency coil 30 and a low frequency coil 40 of theoscillation coil 50, and an electromagnetic wave including a highfrequency alternating magnetic field and a low frequency alternatingmagnetic field was generated, to thereby allow magnetic stimulation tobe applied for 30 minutes to the MB8 cells within each culture well ofthe culture plate 60, and then to be stopped for 30 minutes duringincubation thereof, and finally re-applied for 30 minutes.

At this time, the frequency of the high frequency electric currentapplied to the high frequency coil 30 were incrementally changed at eachexperimental unit within the range of 20 MHz to 3000 MHz, and a highfrequency alternating magnetic field at different high frequencies forthe production of promotion was applied to the cells. Whereas, thefrequency of the low frequency electric current applied to the lowfrequency coil 40 was maintained at 2.0 kHz, and a low frequencyalternating magnetic field at a constant frequency (2.0 kHz) was appliedto the cells. Accordingly, the influence of the low frequencyalternating magnetic field was eliminated to allow the correlationbetween the frequency of the high frequency alternating magnetic fieldand the productivity of the neurotrophic factor group in the MB8 cellsto be experimentally tested. Moreover, regardless of the frequency ofthe high frequency alternating magnetic field, both the high frequencyalternating magnetic field and the low frequency alternating magneticfield were intermittently output at 7.81 Hz, as shown in theabovementioned FIG. 4. Furthermore, when the magnetic field intensity(magnetic flux density) of the center portion of the above-mentionedoscillation coil 50 during magnetic stimulation was measured, themagnetic field intensity of the 83.3 MHz high frequency electromagneticwave was 1.26 μT, and the magnetic field intensity of the low frequencyelectromagnetic wave was 13 μT.

(3) The Cultivation of MB8 Cells after Magnetic Stimulation, and theProduction of the Neurotrophic Factor Group

Each of the MB8 cells (the cells producing the neurotrophic factorgroup) of the magnetically stimulated group that received the magneticstimulation at each frequency of the abovementioned (2) was incubatedfor 3 hours at 37° C. using the abovementioned culture plate. During thecultivation, the MB8 cells produced a neurotrophic factor group in theamount according to each frequency at the time the abovementionedmagnetic stimulation is received, and released it extracellularly.Furthermore, the MB8 cells of the unstimulated group in which themagnetic stimulation of the abovementioned (2) was not implemented werealso cultured under similar conditions to those of the magneticallystimulated group.

(4) Provision of Neurotrophic Factor Group to PC12 Cells, and NeuriteOutgrowth

After the cultivation of the abovementioned (3), the culture medium ofeach of the MB8 cells (including the neurotrophic factor group producedby the MB8 cells) of the magnetically stimulated group was aspirated,filtered with a microfilter, and then each post-filtered culture mediumwas added to the PC12 cells. Afterwards, each of the PC12 cells wascultured at 37° C. for 24 hours. During the cultivation, the neurites ofeach of the PC12 cells were formed and outgrown, depending on the amountof the neurotrophic factor group present within the culture medium.Furthermore, the PC12 cells to which the culture medium of the MB8 cellsof the unstimulated group was added were also cultured in a similarmanner.

(5) Determination of Neurite Outgrowth in PC12 Cells

Each of the cultured PC12 cells after the cultivation of theabovementioned (4) was observed with a microscope, and cells in whichthe length of the neurites was outgrown by at least one cell length weredetermined as positive cells. Three hundred PC12 cells were observed foreach cell group that was magnetically stimulated at each frequency, andthe number of positive cells was recorded. In addition, the PC12 cellsof the unstimulated group were also similarly determined, and the numberof positive cells was recorded.

Then, based on the below-mentioned formula, the effectiveness of themagnetic stimulation at each of the abovementioned frequencies wasdetermined. This effectiveness provides an indication of the degree ofneurite outgrowth of the PC12 cells of the magnetically stimulated groupas compared to that of the PC12 cells of the unstimulated group, i.e.,an indication of the degree of neurite outgrowth of the PC12 cells viamagnetic stimulation (magnetic treatment effect) at each frequency.Specifically, the higher the effectiveness, the greater the amount ofthe neurotrophic factor group produced (high degree of neurotrophicfactor group productivity) within the MB8 cells via magneticstimulation, which indicates that the neurites of the PC12 cells areoutgrown (high degree of neurite outgrowth) via the neurotrophic factorgroup. This indicates that, by supplying a high concentration of theneurotrophic factor group, the repair, growth, differentiation, orproliferation of central nervous system cells or craniospinal nervoussystem cells weakened and the like as a result of a disease waspromoted, to thereby provide a highly effective magnetic treatment for adisease.(Effectiveness)=(Number of positive cells of the magnetically stimulatedgroup)/(Number of positive cells of the unstimulated group)

In addition, after the neurite outgrowth effect described above wasobtained for each experimental unit, these effects were totaled for eachsame frequency, and the average effectiveness at neurite outgrowth(number of times that of the unstimulated group) was determined.

Moreover, the present Experiment 1 was conducted a total of 2,173 timesat all frequencies. When the experimental data from each of theseexperiments were totaled, the data indicating that the neurite outgrowthmight be related to a factor other than magnetic stimulation (e.g., poorcell culture, specific experimental data that deviated considerably fromother experimental data for the same frequency) were eliminated from thetotal. On each day of the present Experiment 1, experiments relating tothe confirmation of the neurite outgrowth of the unstimulated group, theconfirmation of the neurite outgrowth at the time of magneticstimulation at 135 MHz, and the confirmation of the neurite outgrowth atthe time of magnetic stimulation by any one of the frequencies employedin the experiment of the previous day were conducted again, to ensurethe accuracy thereof.

Furthermore, the PC12 cells employed in the abovementioned Experiment 1demonstrate a property whereby differentiation from adrenal cells intoneurocytes is initiated by the action of the nerve growth factor and thelike, and the differentiation of these PC12 cells into neurocytes can beeasily determined based on the neurite outgrowth. The reaction producingthe neurotrophic factor group has several mechanisms, and each reactionthereof is carried out via the interaction of several reactions (cascadereaction). In the present Experiment 1, rather than investigating eachof these mechanisms and reactions, the degree of occurrence of neuriteoutgrowth phenomena that are critical to a living subject and foroverall consequent nerve function was studied. Specifically, even if theabove-mentioned individual reactions relating to the production ofneurotrophic factor group are promoted, since it is thought that thevalue as a treatment would be low if there was no neurite outgrowth as aconsequence, the degree of neurite outgrowth at each frequency wasmeasured as an indicator showing the magnetic treatment effecttherefrom.

As described above, the experimental conditions and experimentalprocedures of Experiment 1 were described. The experimental results ofthe Experiment 1 are shown in Table 1 and FIG. 8. Furthermore, the graphof FIG. 8 illustrates an approximate curve, by plotting the experimentaldata of the average effectiveness (number of times that of theunstimulated group) shown in Table 1 at each frequency (MHz).

TABLE 1

Moreover, the meaning of each parameter of the above Table 1 and FIG. 8is as follows:

“Frequency (MHz)” is a frequency of a high frequency electromagneticwave generated by the abovementioned magnetic stimulation device, i.e.,a frequency of the high frequency alternating magnetic field applied tothe MB8 cells; and “Effectiveness” is the value of the effectiveness ofthe magnetically stimulated group divided by the effectiveness of theunstimulated group and averaged for each frequency, and indicates howmany number of times the neurite outgrowth effect of the magneticallystimulated group via the high frequency alternating magnetic field ofeach frequency is, compared to that of the unstimulated group.

As shown in Table 1 and FIG. 8, when the frequency of the high frequencyalternating magnetic field applied to the MB8 cells (the cells that aretarget of the magnetic stimulation) is within the range of 120 MHz to160 MHz (the first most preferable range), the effectiveness on theneurite outgrowth of the PC12 cells via the neurotrophic factor group isat least 3.5 times that of the unstimulated group, which is extremelyhigh, especially, when the abovementioned frequency is 140 MHz to 160MHz, the effectiveness is at the highest peak of approximately 3.6 timesthat of the unstimulated group. Accordingly, when magnetic stimulationof the high frequency alternating magnetic field within theabovementioned first most preferable range is applied, the outgrowth ofPC12 cell neurites that can be promoted is at least 3.5 times that of acase in which no magnetic stimulation is applied, and as such, anextremely remarkable magnetic treatment effect is exerted.

Furthermore, when the frequency of the high frequency alternatingmagnetic field applied to the MB8 cells (the cells that are the targetof the magnetic stimulation) is within the range of 100 MHz to 160 MHz(the second most preferable range), the effectiveness of the neuriteoutgrowth of the PC12 cells via the neurotrophic factor group is atleast 3.0 times that of the unstimulated group, which is extremely high.Accordingly, when magnetic stimulation of the high frequency alternatingmagnetic field within the abovementioned second most preferable range isapplied, the outgrowth of the PC12 cell neurites that can be promoted isat least 3.0 times that of a case in which no magnetic stimulation isapplied, and as such, an extremely remarkable magnetic treatment effectis exerted.

In addition, when the frequency of the high frequency alternatingmagnetic field is within the range of 60 MHz to 180 MHz, 280 MHz to 300MHz, 450 MHz to 550 MHz, or 900 MHz to 950 MHz (the third mostpreferable range), the effectiveness of the neurite outgrowth of PC12cells via the neurotrophic factor group is at least 2.5 times that ofthe unstimulated group, which is very high. Accordingly, when magneticstimulation of the high frequency alternating magnetic field within theabovementioned third most preferable range is applied, the outgrowth ofthe PC12 cell neurites that can be promoted is at least 2.5 times thatof a case in which no magnetic stimulation is applied, and as such, anextremely remarkable magnetic treatment effect is exerted.

Moreover, when the frequency of the high frequency alternating magneticfield is within the range of 20 MHz to 180 MHz, 280 MHz to 600 MHz, or700 MHz to 1000 MHz (the fourth most preferable range), theeffectiveness of the neurite outgrowth of PC12 cells via theneurotrophic factor group is at least 2.0 times that of the unstimulatedgroup. Accordingly, when magnetic stimulation of a high frequencyalternating magnetic field within the abovementioned fourth mostpreferable range is applied, the outgrowth of PC12 cell neurites thatcan be promoted is at least 2.0 times that of a case in which nomagnetic stimulation is applied, and as such, an extremely remarkablemagnetic treatment effect is exerted.

Furthermore, the neurotrophic factor group that is produced in vivo isreleased from the cells producing neurotrophic factor group, and passesthrough intercellular spaces to reach the site requiring theneurotrophic factor group (the site that is the target of treatment).During this passage thereof, although a molecule of the neurotrophicfactor group passes therebetween, liquid components are reduced (thisphenomenon is referred to as “bioconcentration”). Accordingly, when theneurotrophic factor group reaches the site where it is required, sinceit is more concentrated than the concentration at the time of theproduction, the magnetic treatment effect is further intensified.

According to the results of Experiment 1 described above, from theperspective of the magnetic treatment effect, it is desirable that thehigh frequency for the promotion of production of the high frequencyalternating magnetic field applied to the cells producing theneurotrophic factor group of the MB8 cells and the like (the cells thatare the target of magnetic stimulation) is preferably within theabovementioned fourth most preferable range, more preferably within thethird most preferable range, even more preferably within the second mostpreferable range, and most preferably within the first most preferablerange. Specifically, by applying a high frequency alternating magneticfield with a frequency in such a range, the production of theneurotrophic factor group via the abovementioned cells producingneurotrophic factor group is significantly promoted, so that a highconcentration of neurotrophic factor group is supplied to the centralnervous system cells or craniospinal nervous system cells existing inthe periphery thereof and the like, and the outgrowth of the neurites ofthese cells can be increased by at least 2.0, 2.5, 3.0, or 3.5 timesthat of the unstimulated group. Accordingly, the central nervous systemcells or craniospinal nervous system cells that are weakened, damaged,or reduced as a result of a brain disease and the like can be repaired,grown, differentiated, or proliferated, and the brain disease and thelike can be suitably treated or prevented, to thereby provide a highlyefficient magnetic treatment.

Experiment 2

Next, Experiment 2, which was conducted in order to verify thatintracellular mRNA is increased and that the synthesis of theneurotrophic factor group is promoted via the magnetic stimulation bythe abovementioned magnetic treatment device 10, will be described. InExperiment 2, an increase in the expression of intracellular mRNA aftermagnetic stimulation at 135 MHz was verified via a reversetranscriptase-polymerase chain reaction method (RT-PCR method)

First, the experimental conditions of Experiment 2 and the procedures(1) to (7) thereof will be described.

(1) Cultivation of MB8 Cells

“MB8 cells”, which are glial lineage cells, were employed as the cellsproducing the neurotrophic factor group (the cells that were the targetof magnetic stimulation). The cultivation of these MB8 cells wasconducted in a similar manner to the cell cultivation of theabove-mentioned Experiment 1.

(2) Magnetic Stimulation of MB8 Cells

Using a magnetic stimulation device similar to that of theabovementioned Experiment 1 (refer to FIG. 7), a high frequencyalternating magnetic field (135 MHz) and a low frequency alternatingmagnetic field (2.0 kHz) were irradiated on the MB8 cells, and magneticstimulation was applied thereto for 20 minutes.

(3) Cultivation of MB8 Cells after Magnetic Stimulation

The MB8 cells that received the magnetic stimulation of theabovementioned (2) were incubated for 3 hours at 37° C., and mRNAproduction was promoted inside the MB8 cells.

(4) Extraction of RNA

After the abovementioned cultivation, the culture medium was discardedand an RNA extraction liquid (ISOGEN) was added. Next, the MB8 cellswere crushed by a homogenizer, and allowed to stand for 5 minutes atroom temperature. Afterwards, chloroform was added to this suspension,it was allowed to stand for 10 minutes at room temperature, and thencentrifuged at 12000×g for 15 minutes at 4° C. After that, thesupernatant was collected, the same amount of isopropanol as thesupernatant was added thereto, it was allowed to stand for 10 minutes atroom temperature, and then centrifuged again at 12000×g for 15 minutesat 4° C. Then, it was rinsed by adding 1 ml of 70% ethanol to theprecipitate obtained therefrom, and centrifuged at 12000×g for 5 minutesat 4° C. Thereafter, the precipitate was vacuum dried for 15 minutes ina desiccator, then sufficiently dissolved by adding a DEPC treatedTris-HCl/EDTA solution, to obtain an RNA solution.

(5) Amplification of RNA Via RT-PCR Method

The abovementioned RNA solution, a 10 μM primer, and ultra-pure waterwere placed in a PCR tube, and reacted for 2 minutes at 72° C. Next, 10mM of deoxynucleotide triphosphate solution (dNTP), 100 mM ofdithiothreitol solution (DTT), and 200 unit/μl of reverse transcriptasesolution were added, and a reverse transcription reaction was conductedfor 60 minutes at 42° C. Afterwards, a Tris-HCl/EDTA solution was added,heat treatment was conducted for 7 minutes at 72° C., and a singlestrand cDNA solution was obtained. Ultra-pure water, PCR buffer, 25 mMMgCl₂, 2.5 mM dNTP mixture, 10 μM each of two primer types, and Taqpolymerase, were added to this single strand cDNA solution, which wasplaced in a PCR tube and reacted for 3 minutes at 94° C. Then,denaturation was performed for 30 seconds at 94° C., annealing wasperformed for 1 minute at 45° C., and chain elongation was performed for45 seconds at 72° C., and the abovementioned reactions, as a singlecycle, were repeated for 40 minutes. After that, reaction at 72° C. for5 minutes was conducted, and chain elongation reaction was concluded.

(6) Separation and Detection of RNA (Electrophoresis)

After the abovementioned chain elongation reaction was completed, aloading buffer was added, and RNA was separated by size viaelectrophoresis on 2% (w/v) agarose gel containing ethidium bromide.

(7) Quantitative Determination of RNA

After the abovementioned electrophoresis, the separated RNA wasfluoresced, translated into an image by a “Molecular Imaging FX(manufactured by Bio-Rad Laboratories, Inc.)”, and mRNA was quantifiedusing “Image J” software.

In addition, the increased expression of the intracellular mRNA of theMB8 cells of the unstimulated group in which magnetic stimulation wasnot applied was also quantified in a similar manner to that of the cellsof the abovementioned magnetically stimulated group. Thus, the degree ofincrease in mRNA (the number of times that of the unstimulated group)was determined by dividing the amount of mRNA of the magneticallystimulated group by the amount of mRNA of the unstimulated group. Thedegree of increase in mRNA was determined two times each for BDNF mRNAand for NGF mRNA.

As described above, the experimental conditions and experimentalprocedures of Experiment 2 were described. Then, the experimentalresults of the abovementioned Experiment 2 will be described. Theseexperimental results are shown in FIG. 9.

As shown in FIG. 9, with regard to BDNF mRNA, the cells of themagnetically stimulated group showed expression that was 2.78 times thatof the unstimulated group in the first experiment, and 2.06 times (anaverage of 2.42 times) that of the unstimulated group in the secondexperiment (i.e., increase in BDNF mRNA). Furthermore, with regard toNGF mRNA, the cells of the magnetically stimulated group showedexpression that was 2.20 times that of the unstimulated group in thefirst experiment, and 1.52 times (an average of 1.86 times) that of theunstimulated group in the second experiment (i.e., increase in NGFmRNA).

According to these experimental results, it was verified that, viamagnetic stimulation, using the above-mentioned magnetic treatmentdevice 10, with a high frequency alternating magnetic field of 135 MHz,mRNA for producing BNDF or NGF within the cells producing theneurotrophic factor group was significantly increased when compared tothe unstimulated group. Thus, by applying magnetic stimulation via theabovementioned magnetic treatment device 10, a large amount of BDNF, orNGF neurotrophic factor or the like is synthesized via the increase inmRNA within the cells producing the neurotrophic factor group, and it isreleased extracellularly.

Experiment 3

Hereinafter, Experiment 3, which was conducted in order to verify theintracellular occurrence of exocytosis via magnetic stimulation by theabovementioned magnetic treatment device 10, will be described.

Exocytosis occurs after an increase in the intracellular concentrationof calcium ions, and begins and ends within a few minutes. Whereas, thetime required for mRNA to be increased via magnetic stimulation, totransmit the neurotrophic factor group production instructions to theorgan in charge of production, and to produce the neurotrophic factorgroup (synthesis and extracellular release thereof) is approximately twohours. By utilizing the time difference between both of these, thepresence or absence of a process involving exocytosis (S20→S30 of theabove FIG. 6) and a process involving mRNA (S40 to S44), with regard tothe intracellular production of neurotrophic factor group could beconfirmed.

Therefore, similarly to the abovementioned Experiment 1, the presentExperiment 3 determined: (1) cell culture; (2) magnetic stimulation; (4)cultivation of PC12 cells; and (5) neurite outgrowth. However, in (3)the cultivation of MB8 cells after magnetic stimulation of Experiment 3,the times that the MB8 cells were left to stand after magneticstimulation (time of producing neurotrophic factor group) were set at 10minutes (10 minute standing group) and 3 hours (3 hour standing group).Although exocytosis occurred in the MB8 cells in the short time span of10 minutes, the synthesis of the neurotrophic factor group accompanyingan increase in mRNA did not occur. Thus, the results of the aboveExperiment 3 prove that when the neurites of the PC12 cells of the 10minute standing group were outgrown, exocytosis occurred in the MB8cells, and the neurotrophic factor group was released.

First, the experimental conditions of Experiment 3 and the procedures(1) to (5) thereof will be described.

(1) Cell Culture

Similarly to (1) of the abovementioned Experiment 1, the MB8 cells thatwere the target cells of magnetic stimulation (cells producing theneurotrophic factor) and the PC12 cells that were the cells used forconfirmation of neurite outgrowth were both cultured for 24 hours.

(2) Magnetic Stimulation

Using a magnetic stimulation device similar to that of theabovementioned Experiment 1 (refer to FIG. 7), magnetic stimulation wasapplied to the MB8 cells within the abovementioned culture plate for 30minutes, followed by 30 minutes of cultivation (no magnetic stimulationwas applied during the cultivation), and then the re-application of themagnetic stimulation for 30 minutes. At such a time, the frequency ofthe high frequency alternating magnetic field that was applied to theMB8 cells was 120 MHz.

(3) Allowing the Cells to Stand after Magnetic Stimulation

The culture plate of the abovementioned post-magnetic stimulation MB8cells of the 10 minute standing group was allowed to stand for 10minutes in a carbon dioxide culture apparatus with a 5% carbon dioxideconcentration at 37° C., while the 3 hour standing group was allowed tostand for 3 hours. Accordingly, in the 10 minute standing group, theneurotrophic factor group was released into the culture medium viaexocytosis. Whereas, in the 3 hour standing group, the release of theneurotrophic factor group via exocytosis and the production and releaseof the neurotrophic factor group via other processes (synthesis andrelease via an increase in mRNA) are both occurred. After the productionof this neurotrophic factor group, the entire culture medium of the MB8cells of the abovementioned 10 minute standing group and 3 hour standinggroup was aspirated, the culture media thereof were filtered with amicrofilter, and a culture medium to be added to the PC12 cells wasobtained.

(4) Cultivation of PC12 Cells

The culture media of the PC12 cells cultured via the abovementioned (1)were removed by aspiration, and the culture medium of the 10 minutestanding group and the culture medium of the 3 hour standing groupobtained via (3) were individually added. After that, the PC12 cellswith each culture medium added thereto were individually cultured for 24hours in a carbon dioxide culture apparatus with a 5% carbon dioxideconcentration at 37° C.

(5) Determination of Neurite Outgrowth in PC12 Cells

Twenty-four hours after the time at which each of the culture mediaobtained by (3) was individually added to the PC12 cells in theabovementioned (4), similarly to the abovementioned Experiment 1, thenumber of positive PC12 cells in which neurites were outgrown wascounted, and the neurite outgrowth ratio (ratio of positive cells) wascalculated.

As described above, the experimental conditions of Experiment 3 and theprocedures thereof were described. Then, the experimental results of theabove Experiment 3 will be described with reference to Table 2.

TABLE 2 Neurite Outgrowth Sample Ratio (%) Unstimulated Group 7.4%Post-Magnetic Stimulation 15.5% 10 Minute Standing Group Post-MagneticStimulation 27.9% 3 Hour Standing Group

As shown in Table 2, the neurite outgrowth ratio of the PC12 cells was7.4% in the unstimulated group (reference group) in which no magneticstimulation is applied. Whereas, the post-magnetic stimulation 10 minutestanding group had an outgrowth ratio of 15.5%, which was 2.1 times thatof the unstimulated group. Accordingly, exocytosis occurred in thepost-magnetic stimulation MB8 cells within a short time period of 10minutes, so that the production of the neurotrophic factor group couldbe verified.

Moreover, the post-magnetic stimulation 3 hour standing group had anoutgrowth ratio of 27.9%, which was 3.8 times that of the unstimulatedgroup and 1.8 times that of the 10 minute standing group. Accordingly,the fact that the production of the neurotrophic factor group occurredin the post-magnetic stimulation MB8 cells via a process other thanexocytosis could be verified.

Experiment 4

Next, Experiment 4, which was conducted in order to verify that theintracellular concentration of calcium ions was increased via magneticstimulation by the above-mentioned magnetic treatment device 10, will bedescribed. In the Experiment 4, cells were collected from each region ofa bovine brain, magnetic stimulation was applied to each of these cellsat 83.3 MHz, 2 kHz, and 7.8 Hz, and the regions of cells in the brain inwhich an increase in the intracellular concentration of calcium ions wasconfirmed and the positive reaction thereof were verified.

First, the experimental conditions of the present Experiment 4 and theprocedures (1) to (5) thereof will be described.

(1) Collection and Cultivation of Cells

Brain slices were anatomically collected from each region of the bovinebrain (frontal lobe region of the cerebral cortex, temporal lobe regionof the cerebral cortex, cerebellum and medulla oblongata regions, andhippocampus), and the cells thereof were primarily cultured as testcells in accordance with a typical brain cell culture method.

(2) Loading of Calcium Fluorescent Indicator

A Fluo-3 calcium fluorescent indicator (Manufactured by DojindoMolecular Technologies, Inc.) was employed to measure the intracellularconcentration of calcium ions. The calcium fluorescent probe (Fluo-3)was added to the above-mentioned test cells cultured in a glass-basedish to bring the final concentration thereof to 4 μM, then loaded for30 minutes at 37° C., washed 3 times with the standard solution, andmeasurements were obtained. The composition of this reference solutionwas 135 mM NaCl, 2.8 mM KCl, 1.8 mM MgCl₂, 10 mM D-glucose, 10 mM HEPES(pH=7.3).

(3) Magnetic Stimulation

A glass-base dish into which cells were placed after the loading of theabovementioned (2) was mounted on an inverted microscope. Theoscillation coil 50 shown in the abovementioned FIG. 7 was mounted onthe lid of this glass-base dish, and magnetic stimulation was applied tothe cells for 10 minutes. During this magnetic stimulation, the 83.3 MHzhigh frequency alternating magnetic field and the 2 kHz low frequencyalternating magnetic field were intermittently irradiated at 7.8 Hz ontothe cells, as shown in FIG. 4.

(4) Measurement of the Distribution of Intracellular Fluorescence

Using an inverted microscope, the magnetically stimulated cells thatwere loaded and stained with the above-mentioned fluorescent dye wereobserved at room temperature (25° C.). A 20- to 40-fold objective lenswas used to allow the fluorescent intensity of at least ten cells to bemeasured simultaneously. The fluorescence from the irradiation ofexcited light was detected with a digital CCD camera (Product name: HiSCA, manufactured by Hamamatsu Photonics K.K.). The fluorescentintensity ratio of the cells was analyzed with a time plus system(Product name: AQUACOSMOS, manufactured by Hamamatsu Photonics K.K.).

Before applying the magnetic stimulation of the abovementioned (3), achange in the fluorescent intensity of the cells that was within 1±0.05was confirmed for at least 5 minutes. Next, a change in theintracellular concentration of calcium ions was observed for a period of30 minutes, after the magnetic stimulation of the abovementioned (3) wasapplied for 10 minutes. Then, 600 mM potassium chloride of only theamount of one-tenth of the volume of liquid in the glass-base dish.Before, during, and after the magnetic stimulation, and after theaddition of potassium chloride, the series of reactions was continued,and intracellular fluorescence intensity distribution was measured andobserved.

(5) Determination of Experimental Success or Failure

Regarding the addition of the abovementioned potassium chloride, only ina case where the cells showed a dramatic increase in the concentrationof calcium ions, the experimental data of such cells were employed.Regarding the addition of potassium chloride, since the cells notshowing a normal reaction in which there was a dramatic increase in theconcentration of calcium ions failed to demonstrate a normal calciumresponse, the experimental data thereof were not employed.

(6) Determination

After the magnetic stimulation, in cases where there was at least onecell in which the intracellular fluorescence intensity increased atleast 10% more than before the magnetic stimulation, it was judged as apositive reaction (i.e., the intracellular concentration of calcium ionswas increased via the magnetic stimulation).

As described above, the experimental conditions and the experimentalprocedures of Experiment 4 were described. Next, the experimentalresults of Experiment 4 will be described with reference to Table 3.

TABLE 3 Positive Reaction Rate Regions of Cells Inside Brain (n: Numberof Samples) Frontal Lobe Region of Cerebral Cortex 57.5% (n = 33)Temporal Lobe Region of Cerebral Cortex 45.4% (n = 22) Cerebellum andMedulla Oblongata Regions 15.3% (n = 13) Hippocampus  5.2% (n = 19)

As shown in Table 3, the increase in the intracellular concentration ofcalcium ions (positive reaction rate) via magnetic stimulation wasrelatively high in the frontal lobe region of the cerebral cortex at57.5%, and the temporal lobe region of the cerebral cortex at 45.4%.Accordingly the cells of these regions in the brain were verified ashaving an increased intracellular concentration of calcium ions viamagnetic stimulation. At such a time, the increase in the intracellularconcentration of calcium ions via magnetic stimulation means that theexocytosis of the above-mentioned cells can be induced and the releaseof the neurotrophic factor group can be promoted.

On the other hand, the increase in the intracellular concentration ofcalcium ions (positive reaction rate) was relatively low in thecerebellum and medulla oblongata regions at 15.3%, and in thehippocampus at 5.2%. Accordingly, it was clear that the increase in theintracellular concentration of calcium ions via magnetic stimulationdiffered depending on the regions within the brain from which cells werecollected.

Experiment 5

Next, Experiment 5, which was conducted in order to verify theproduction of substance (i.e., a neurotrophic factor-like substance)that demonstrates a neurite outgrowth effect other than the neurotrophicfactor via magnetic stimulation by the abovementioned magnetic treatmentdevice 10, will be described.

In addition to the abovementioned various types of neurotrophic factors,adenosine, adenosine monophosphate (AMP), a manganese ion, genipin,lysophosphatidylethanolamine, Rho-kinase and the like are known assubstances that demonstrate an effect whereby the neurites of the neuralcells are outgrown. The present experiment 5 was conducted to confirmwhether a substance (a neurotrophic factor-like substance) thatdemonstrates a neurite outgrowth effect other than the neurotrophicfactor, and the neurotrophic factor are produced via magneticstimulation on MB8 cells.

Since neurotrophic factor is protein, it is easily denatured via theapplication of heat, and the neurite outgrowth effect is lost. Aproteinaceous component and a non-proteinaceous component are present inthe neurotrophic factor-like substance. The non-proteinaceous componentdoes not lose its neurite outgrowth effect via the application of heat.Thus, in the present Experiment 5, a post-magnetically stimulatedculture medium of the MB8 cells to which heat was applied and one towhich heat was not applied were added to the PC12 cells, individually,and the degree of neurite outgrowth of the PC12 cells to which theheated culture medium (heated group) was added and the degree of neuriteoutgrowth of the PC12 cells to which the unheated culture medium(unheated group) was added were compared to confirm the presence of theneurotrophic factor or the neurotrophic factor-like substance. If theneurites of the PC12 cells to which the heated culture medium was addedare outgrown, the existence of the neurotrophic factor-like substancecan be verified thereby.

(1) Cultivation of MB8 Cells

The culture medium of MB8 cells cultured in a similar manner to that ofthe abovementioned Experiment 1 was sampled, and 400 μl of serum-freeRPMI was added thereto.

(2) Magnetic Stimulation on MB8 Cells

Using a magnetic stimulation device similar to that of theabovementioned Experiment 1 (refer to FIG. 7), magnetic stimulation by ahigh frequency alternating magnetic field (135 MHz) was applied to theabovementioned MB8 cells for 30 minutes, which were then incubated for30 minutes under a 5% carbon dioxide concentration at 37° C. (nomagnetic stimulation was applied during the cultivation), and afterwardsagain subjected to 30 minutes of magnetic stimulation.

(3) Cultivation of MB8 Cells after Magnetic Stimulation, and Productionof Neurotrophic Factor Group

Each of the MB8 cells (cells producing the neurotrophic factor group) ofthe magnetically stimulated group receiving the magnetic stimulation ateach frequency of the abovementioned (2) was cultured for 3 hours undera 5% carbon dioxide concentration at 37° C.

(4) Heating of Culture Medium

After the cultivation of the abovementioned (3), the entire volume ofthe culture medium of the MB8 cells (including the neurotrophic factorgroup produced by the MB8 cells) was collected in a microtube, andheated for 2 minutes in 90° C. oil bath. After 2 minutes of thisheating, the microtube was taken out and rapidly cooled in an ice waterfor 1 minute. The neurite outgrowth effect of the neurotrophic factor(protein) included in the culture medium was lost via this heatingprocess.

(5) Filtration of Culture Medium

Fetal bovine serum (FBS) was added to the culture medium obtained viathe abovementioned (4) to bring the concentration up to 1%, the culturemedium was filtered with a filter, and the coagulum was eliminated.

(6) Supply of Neurotrophic Factor Group to PC12 Cells, and NeuriteOutgrowth

The culture medium of the PC12 cells cultured in a similar manner tothat of the abovementioned Experiment 1 was aspirated, and the culturemedium filtered via (5) was added to the PC12 cells. Thereafter, thePC12 cells were cultured for 24 hours under a 5% carbon dioxideconcentration at 37° C.

As described above, a heated group sample, in which a heated culturemedium of the MB8 cells was added to the PC12 cells, was formed fromamong the magnetically stimulated group. Moreover, an unstimulated groupsample in which magnetic stimulation was not applied to the MB8 cells,and an unheated group sample from the magnetically stimulated group inwhich the heating of the abovementioned (4) was not applied, were formedin a similar manner to that of the above-mentioned Experiment 1.

(7) Determination of Neurite Outgrowth in PC12 Cells

Regarding each of the abovementioned unstimulated group, heated group,and unheated group, the number of positive PC12 cells in which neuriteswere outgrown was counted in a similar manner to that employed by theabovementioned Experiment 1, and the neurite outgrowth ratio (ratio ofpositive cells) thereof was calculated.

As described above, the experimental conditions and the experimentalprocedures of the abovementioned Experiment 5 were described. Then, theexperimental results of the Experiment 5 will be described withreference to FIG. 10. FIG. 10 is a graph showing the neurite outgrowthratio of the unstimulated group, the unheated group, and the heatedgroup.

As shown in FIG. 10, the neurite outgrowth ratio of the unstimulatedgroup, the unheated group, and the heated group were 1.1%, 34.4%, and21.3%, respectively. First, the unstimulated group and the heated groupwere compared. According to the abovementioned experimental results, itwas confirmed that even the heated group altered via the heating of theneurotrophic factor had a neurite outgrowth ratio that was approximately1.9 times higher (=21.1%/11.1%) than that of the unstimulated group.Moreover, PC12 cells in which the morphology thereof had changed to thatof a neural cell were also observed in the heated group. According tothese experimental results, the presence of a substance, other than aneurotrophic factor, demonstrating the neurite outgrowth effect wasclearly verified. Therefore, the MB8 cells produced, via magneticstimulation, not only a neurotrophic factor which is protein(thermolabile component) but also a substance demonstrating a neuriteoutgrowth effect other than these (i.e., a neurotrophic factor-likesubstance: including thermostable component), and as such, the neuritesof the PC12 cells were said to be outgrown via the neurotrophicfactor-like substance.

Next, when the unheated group and heated group were compared, theoutgrowth ratio of the heated group was approximately 62% that of theunheated group (=21.3%/34.4%). Accordingly, when the neurite outgrowtheffect via magnetic stimulation (the outgrowth ratio of the unheatedgroup) was 100%, the neurite outgrowth effect was reduced approximately38% by heating, as a result of the thermolabile component (i.e.,neurotrophic factor and proteinaceous neurotrophic factor-likesubstance). That is to say, the neurite outgrowth effect via a component(the thermostable component of the neurotrophic factor-like substance)that has not been denatured by heat is approximately 62%. Accordingly,based on the abovementioned experimental results, the presence of anon-proteinaceous component (i.e., neurotrophic factor-like substance)with the proteinaceous component (i.e., neurotrophic factor andproteinaceous neurotrophic factor-like substance) excluded therefrom wasverified. However, since not only was the neurotrophic factor, but alsoa proteinaceous substance other than a neurotrophic factor and aproteolytic enzyme that is a neurite formation inhibitor were denaturedby heating, it can not be said for certain that all of the neuriteoutgrowth effects of the heated group resulted from thenon-proteinaceous component. Since there is a variety of types ofsubstances other than the proteinaceous neurotrophic factor andproteolytic enzymes, it is extremely difficult to measure these resultsand calculate the effects thereof.

According to the abovementioned experimental results, the MB8 cellsproduced not only, via magnetic stimulation, the neurotrophic factorprotein but also the neurotrophic factor-like substance, and theneurites of the PC12 cells can be outgrown by the action of thisneurotrophic factor-like substance. Accordingly, via the reception ofmagnetic stimulation, cells producing a neurotrophic factor of a glialcell and the like were verified as producing, in addition toneurotrophic factor, the neurotrophic factor-like substancedemonstrating the neurite outgrowth effect on the central nervous systemcells or craniospinal nervous system cells.

As described above, according to an embodiment of the present invention,Experiments 1-5 were described. According to the experimental resultsmentioned above, it was verified that employing the magnetic treatmentdevice 10 of the present embodiment to apply a high frequencyalternating magnetic field of a desired frequency to the cells of anaffected area, allows the intracellular concentration of calcium ions tobe increased so as to induce the exocytosis of the neurotrophic factorgroup, and mRNA of the intracellular neurotrophic factor group to beincreased so as to promote the synthesis and release of theintracellular neurotrophic group. Moreover, it was verified that bypromoting the production of the neurotrophic factor group in such amanner, the production of the neurotrophic factor group of the centralnervous system cells or craniospinal nervous system cells is promoted,and the repair, growth, differentiation, or proliferation of these cellsis promoted, so that various diseases such as a brain disease can betreated.

Although the preferred embodiments of the present invention have beenexplained above with reference to the appended drawings, the presentinvention is not specifically limited to such an example. Thus, so longas one is skilled in the art, it is clear that various alternativeembodiments or modified embodiments covered within the scope of theappended claims would be readily apparent, and thus it is understoodthat all such alternatives and/or modifications are naturally includedwithin the technical scope of the present invention.

For example, although the high frequency and low frequencyelectromagnetic wave generating means of the abovementioned embodimentinclude a coil, such as the high frequency coil 30 or the low frequencycoil 40 as the antenna emitting the electromagnetic wave, the presentinvention is not specifically limited to such an example. For example,in addition to a loop antenna, such as a coil or the like, the antennaemitting the electromagnetic wave may also be constructed of varioustypes of antennas, such as a rod antenna, a Hertz dipole antenna, ashort antenna, a half-wave dipole antenna, a helical antenna, a monopoleantenna, a rhombic antenna, an array antenna, a horn antenna, aparabolic antenna, or a slot antenna. Moreover, the coil employed as theabovementioned antenna may be configured from a solenoid coil, aHelmholtz antenna, a rotary coil, a split pair coil, a shim coil, or asaddle type coil or the like. Furthermore, the material, the shape, thesize, the number of turns, the presence or absence of a shaft core, andthe location and the like of the high frequency coil 30 and lowfrequency coil 40 are also not specifically limited to the examples ofthe abovementioned embodiment (FIG. 2A and FIG. 2B), and as such, designmodifications are possible where needed.

Moreover, in the abovementioned embodiment, although a circuit structuresuch as that of the control block 20 shown in FIG. 3 is employed as thehigh frequency oscillation means and the low frequency oscillation meansapplying the high frequency electric current or the low frequencyelectric current to the high frequency coil 30 or the low frequency coil40, the present invention is not specifically limited to such anexample. For example, so long as the circuit structure of the controlblock 20 allows for the oscillation of a high frequency wave within thepredetermined range of the high frequency for the promotion ofproduction, various design modifications are possible. For example, thehigh frequency oscillation means 24 that is capable of the oscillationof the abovementioned high frequency wave and the low frequencyoscillation means 25 that is capable of the oscillation of thepredetermined low frequency wave (e.g., 2.0 kHz, and 7.81 Hz) may beincluded without the necessity to include a main control circuitcomposed of a micro-computer and the like.

In addition, although 83.3 MHz or 135 MHz is mainly exemplified as thehigh frequency for the promotion of production of the high frequencyelectromagnetic wave (high frequency alternating magnetic field) in theabove-mentioned embodiments and the Examples, so long as the highfrequency for the promotion of production is a predetermined frequencywithin the range of 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700 MHz to1000 MHz (the fourth most preferable range), the present invention isnot specifically limited to such an example. Furthermore, although 2.0kHz is exemplified as the low frequency for the production of promotionof the low frequency electromagnetic wave, it is not specificallylimited to such an example, and as such, may include a predeterminedfrequency within the range of approximately 2.0±10% kHz, or an arbitraryfrequency within a range other than this one.

Moreover, although the high frequency electromagnetic wave of theabovementioned embodiment is a substantially sinusoidal wave, it is notspecifically limited thereto. For example, a substantially rectangularwave, a saw-tooth wave or the like, may also be included. In addition,although the low frequency electromagnetic wave is a substantiallyrectangular wave, it is not specifically limited thereto. For example, asubstantially sinusoidal wave, a saw-tooth wave or the like, may also beincluded. In addition, although the above-mentioned low frequencyelectromagnetic wave is a substantially rectangular wave of two values,a positive predetermined value and a zero value, it is not specificallylimited to these two values. For example, both values may be positive ornegative, or one value may be positive and the other negative, or thelike.

Furthermore, although the high frequency electromagnetic wave generatingmeans in the abovementioned embodiment intermittently generates a highfrequency electromagnetic wave of a composite frequency of approximately2.0 kHz and approximately 7.81 Hz, the present invention is notspecifically limited thereto. For example, the high frequencyelectromagnetic wave generating means may also intermittently generate ahigh frequency electromagnetic wave of a frequency of eitherapproximately 2.0±10% kHz or approximately 7.81±10% Hz alone, or maycontinuously generate a non-intermittent high frequency electromagneticwave of at least one frequency other than the abovementionedfrequencies.

Moreover, the high frequency electromagnetic wave generating means maynot only completely intermittently generate the high frequencyelectromagnetic wave, but also may generate the high frequencyelectromagnetic wave so that the electromagnetic wave intensity thereofis sinusoidally increased and decreased at the predetermined at leastone frequency (e.g., approximately 2.0±10% kHz and approximately7.81±10% Hz), for example. Since this also allows the intensity of thehigh frequency alternating magnetic field applied to the subject to betreated to be periodically increased and decreased, and the stimulationof the alternating magnetic field to be changed, the magnetic treatmenteffect is enhanced. Furthermore, for example, a low frequencyelectromagnetic wave generated by the low frequency electromagnetic wavegenerating means may be periodically increased and decreased or may beinterrupted in synchronization with the periodic increases and decreasesin the high frequency electromagnetic wave intensity.

In addition, although the low frequency electromagnetic wave generatingmeans of the abovementioned embodiment intermittently generates a lowfrequency electromagnetic wave at a cycle of approximately 7.81 Hz, thepresent invention is not specifically limited thereto. For example, thelow frequency electromagnetic wave generating means may alsointermittently generate a low frequency electromagnetic wave of at leastone frequency other than the above-mentioned frequencies. Furthermore,the low frequency electromagnetic wave generating means may continuouslygenerate a non-intermittent low frequency electromagnetic wave.

Moreover, although the magnetic treatment device 10 of theabovementioned embodiment is constructed to be capable of generatingboth a high frequency electromagnetic wave and a low frequency magneticwave, by including both the high frequency oscillation means 24 and thelow frequency oscillation means 25 therein, the present invention is notspecifically limited thereto. The magnetic treatment device 10 may alsohave a structure for solely generating the abovementioned high frequencyelectromagnetic wave, without including the abovementioned low frequencyoscillation means 25. Furthermore, in addition to the abovementionedhigh frequency oscillation means 24 and/or the abovementioned lowfrequency oscillation means 25, the magnetic treatment device 10 mayadditionally include at least one novel electromagnetic wave generatingmeans (e.g., a separate coil). In addition, the electromagnetic wavegenerated by the additional electromagnetic wave generating means mayalso be an electromagnetic wave of an arbitrary frequency, such as along wave, a medium wave, a short wave, an ultra short wave, amicrowave.

Moreover, aside from a structural element other than that describedabove, the magnetic treatment device 10 may also optionally include avibration generating means for providing vibrations to the subject to betreated; various measuring devices for measuring the frequency orintensity of the electromagnetic wave (alternating magnetic field) to beapplied, room temperature, body temperature, amount of batteryremaining, and the like; a timer device for automatically activating anon/off mode or the like of an operation, by measuring and controllingthe continuous irradiation time of the alternating magnetic field(operation time); a sound generating device, such as a buzzer device fornotifying a user via voice of the end of the scheduled treatment time orpower consumption; an attaching means, such as a belt or adhesive agent,for attaching the main body of the treatment device on the affectedarea; and the like.

Furthermore, although the magnetic treatment device 10 of theabovementioned embodiment is configured to generate a high frequencyelectromagnetic wave of the high frequency for the promotion ofproduction that is selected from the range of 20 MHz to 180 MHz, 280 MHzto 600 MHz, or 700 MHz to 1000 MHz, the present invention is notspecifically limited thereto.

For example, the magnetic treatment device 10 may be configured so as tooscillate a frequency in which an arbitrary frequency within the rangeof the above-mentioned high frequency for the promotion of production isdivided by an arbitrary positive integer (e.g., approximately 75 MHz, 50MHz, 37.5 MHz, 30 MHz, . . . , which are 150 MHz divided by the positiveintegers 2, 3, 4, 5, . . . ), and configured so as to also generate thehigh frequency electromagnetic wave of the abovementioned high frequencyfor the promotion of production using a higher harmonic wave thatadditionally occurs when generating the electromagnetic wave of thisfrequency.

Specifically, in general, when the fundamental harmonic of the highfrequency electromagnetic wave that is generated is not a completelysinusoidal wave, the higher harmonic wave is inevitably generated alsofor a frequency that is an integral multiple of the fundamentalharmonic. FIG. 11 is a graph measuring the distribution of frequencythat is actually generated from the magnetic treatment device 10 whenthe frequency of the high frequency electromagnetic wave generated bythe magnetic treatment device 10 of the above-mentioned embodiment isset at 80 MHz. As shown in FIG. 11, when the magnetic treatment device10 that is set at 80 MHz, the high frequency electromagnetic wave of afrequency (e.g., 160 MHz, 240 MHz, 320 MHz, 400 MHz, 480 MHz, . . . )that is an integral multiple (e.g., 2×, 3×, 4×, . . . ) of 80 MHz isgenerated as the higher harmonic wave.

So long as the frequency of the higher harmonic wave generated in thismanner is within the desired range, for example, 20 MHz to 180 MHz, 280MHz to 600 MHz, or 700 MHz to 1000 MHz (within the fourth mostpreferable range), of the high frequency for the promotion of productionof the abovementioned embodiment, it is thought that the magnetictreatment effect occurs from the application of the abovementionedhigher harmonic wave to the subject to be treated. Accordingly, themagnetic treatment device and the neurotrophic factor productionpromoting device generating the fundamental harmonic as the generatingsource of the abovementioned higher harmonic wave are included withinthe technical scope of the present invention.

In addition, the magnetic treatment device 10 may also generate a highfrequency electromagnetic wave of a high frequency for the promotion ofproduction that is within the abovementioned first to fourth mostpreferable ranges by intermittently generating a high frequencyelectromagnetic wave of a frequency that is higher than that of theabovementioned first to fourth most preferable ranges (greater than 1000MHz) with a frequency that is within the abovementioned first to fourthmost preferable ranges.

Specifically, even if the living cells of the human body and the likereceive the irradiation of an electromagnetic wave of a frequency bandof an excessively high frequency, they may not react to changes in thealternating magnetic field of the abovementioned high frequency. Byutilizing the insensitivity of the above living cells, a high frequencyelectromagnetic wave that is higher than that of the abovementionedfirst to fourth most preferable ranges (e.g., 1 GHz) is generated as acarrier wave, and the above-mentioned carrier wave is output by beingturned on/off at a cycle corresponding to a frequency within the firstto fourth most preferable ranges (e.g., 150 MHz), which is theabove-mentioned frequency for the production of promotion, to therebyallow the living cells to react as if only an electromagnetic wave ofthe high frequency wave for the production of promotion was irradiated.Accordingly, the magnetic treatment device and the neurotrophic factorproduction promoting device that intermittently generate the carrierwave as the generating source of the high frequency wave are includedwithin the technical scope of the present invention.

Moreover, the high frequency for the promotion of production may also beof a fixed value within the abovementioned first to fourth mostpreferable ranges. However, when the high frequency alternating magneticfield of the same high frequency for the production of promotion iscontinuously applied to the cells of an affected area, there is apossibility that the cells of the affected area will become accustomedto the above frequency, and thus the magnetic treatment effect will bereduced. Thus, during the treatment employing the abovementionedmagnetic treatment device 10 (during the irradiation of the highfrequency alternating magnetic field on the affected area), theabovementioned high frequency for the promotion of production may alsobe changed within the abovementioned first to fourth most preferableranges. Accordingly, since high frequency alternating magnetic fields ofdifferent high frequencies for the promotion of production may beapplied to the cells of an affected area during the magnetic treatment,the magnetic stimulation which the cells receive is changed, and themagnetic treatment effect can be enhanced. Moreover, the above-describedchanges in the high frequency for the promotion of production can beachieved by, for example, changing the frequency of the high frequencyelectric current applied to the high frequency coil 30 by the highfrequency oscillation means within the abovementioned range.

In addition, although an example of the neurotrophic factor productionpromoting device utilized as the magnetic treatment device 10 for theapplication of magnetic stimulation to the affected area of a livingbody was described, the present invention is not specifically limited tosuch an example. So long as the neurotrophic factor production promotingdevice of the present invention is a device promoting the production ofthe neurotrophic factor group by the application of magnetic stimulationto cells, various devices, such as a test device applying magneticstimulation to cells isolated from the subject to be treated (e.g.,human body, or animal), for example, may also be applicable.

Furthermore, the neurotrophic factor and neurotrophic factor-likesubstance are not specifically limited to the substances illustrated bythe abovementioned embodiments. So long as the neurotrophic factor andthe neurotrophic factor-like substance of the present invention aresubstances contributing to the repair, growth, differentiation orproliferation of central nervous system cells or craniospinal nervoussystem cells, any currently known substances, as well as any substancesthat may be discovered in the future, may be included therein.

INDUSTRIAL APPLICABILITY

The present invention is applicable to a production promoting device foran intracellular neurotrophic factor group, specifically it isapplicable to a magnetic treatment device for treating aneurodegenerative disorder such as Alzheimer's disease, depression, orthe like.

The invention claimed is:
 1. A device configured to promote theproduction of a neurotrophic factor via application of magneticstimulation to cells, the device comprising: a high frequencyelectromagnetic wave generating means generating a high frequencyelectromagnetic wave of a high frequency for promotion of productionselected from the range of 20 MHz to 180 MHz, 280 MHz to 600 MHz, or 700MHz to 1000 MHz and comprising a high frequency coil that generates thehigh frequency electromagnetic wave, a low frequency electromagneticwave generating means generating a low frequency electromagnetic wavelower than the high frequency electromagnetic wave and comprising a lowfrequency coil that generates the low frequency electromagnetic wave andwherein the low frequency coil is different from the high frequencycoil; and a frequency controlling means that generates the high and lowfrequency electromagnetic waves intermittently and such that the highand low frequency electromagnetic waves are synchronized.
 2. The deviceof claim 1, wherein the high frequency for the promotion of productionis selected from the range of 60 MHz to 180 MHz, 280 MHz to 300 MHz, 450MHz to 550 MHz, or 900 MHz to 950 MHz.
 3. The device of claim 1, whereinthe high frequency for the promotion of production is selected from therange of 100 MHz to 160 MHz.
 4. The device of claim 1, wherein the highfrequency for the promotion of production is selected from the range of120 MHz to 160 MHz.
 5. The device of claim 1, wherein the high frequencyelectromagnetic wave generating means further comprises a high frequencyoscillation means outputting a high frequency electric current; and ahigh frequency antenna generating the high frequency electromagneticwave of the high frequency for the promotion of production viaapplication of a high frequency current from the high frequencyoscillation means.
 6. The device of claim 1, wherein the high frequencyelectromagnetic wave generating means intermittently generates the highfrequency electromagnetic wave, by repeating an on time period in whichthe high frequency electromagnetic wave is generated and an off timeperiod in which the high frequency electromagnetic wave is notgenerated, at a predetermined cycle.
 7. The device of claim 6, whereinthe high frequency electromagnetic wave generating means intermittentlygenerates the high frequency electromagnetic wave, by repeating a firston time period in which the high frequency electromagnetic wave isgenerated and a first off time period in which the high frequencyelectromagnetic wave is not generated, at a cycle corresponding to2.0±10% kHz.
 8. The device of claim 6, wherein the high frequencyelectromagnetic wave generating means intermittently generates the highfrequency electromagnetic wave, by repeating a second on time period inwhich the high frequency electromagnetic wave is generated and a secondoff time period in which the high frequency electromagnetic wave is notgenerated, at a cycle corresponding to 7.8±10% Hz.
 9. The device ofclaim 1, wherein the low frequency electromagnetic wave generating meansfurther comprises a low frequency oscillation means outputting a lowfrequency electric current; and a low frequency antenna generating thelow frequency electromagnetic wave of the low frequency for thepromotion of production via application of a low frequency current fromthe low frequency oscillation means.
 10. The device of claim 9, whereina rise time of the low frequency electric current applied to the lowfrequency antenna is no more than 0.1 μsec.
 11. The device of claim 1,wherein the low frequency electromagnetic wave generating meansintermittently generates the low frequency electromagnetic wave, byrepeating an on time period in which the low frequency electromagneticwave is generated and an off time period in which the low frequencyelectromagnetic wave is not generated, at a predetermined cycle.
 12. Thedevice of claim 11, wherein the low frequency electromagnetic wavegenerating means intermittently generates the low frequencyelectromagnetic wave, by repeating a third on time period in which thelow frequency electromagnetic wave is generated and a third off timeperiod in which the low frequency electromagnetic wave is not generated,at a cycle corresponding to 7.8±10% Hz.
 13. The device of claim 11,wherein the high frequency electromagnetic wave generating meansintermittently generates the high frequency electromagnetic wave, byrepeating an on time period in which the high frequency electromagneticwave is generated and an off time period in which the high frequencyelectromagnetic wave is not generated, at a predetermined cycle, and theon time period of the high frequency electromagnetic wave and the ontime period of the low frequency electromagnetic wave are synchronizedwith each other.
 14. The device of claim 1, wherein the low frequencyelectromagnetic wave generating means generates a low frequencyelectromagnetic wave in the range of 2.0±10% kHz.
 15. A method forpromoting neurotrophic factor production in a cell, the methodcomprising applying magnetic stimulation to the cells with the device ofclaim 1, wherein the magnetic stimulation by the high frequencyalternating magnetic field of the high frequency is configured andsuitable for increasing a concentration of calcium ions within the cellsso that exocytosis of the neurotrophic factor is induced, and themagnetic stimulation increases synthesis of messenger ribonucleic acid(mRNA) of the neurotrophic factor in the cells so that the synthesis andextracellular release of the neurotrophic factor increases.